CA2554703A1 - Differential expression of markers in ovarian cancer - Google Patents
Differential expression of markers in ovarian cancer Download PDFInfo
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- CA2554703A1 CA2554703A1 CA002554703A CA2554703A CA2554703A1 CA 2554703 A1 CA2554703 A1 CA 2554703A1 CA 002554703 A CA002554703 A CA 002554703A CA 2554703 A CA2554703 A CA 2554703A CA 2554703 A1 CA2554703 A1 CA 2554703A1
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Abstract
Novel markers for ovarian cancer that are both sensitive and accurate. These markers are overexpressed and/or differentially expressed in ovarian cancer specifically, as opposed to normal ovarian tissue. The measurement of these markers, alone or in combination, in patient samples provides information that the diagnostician can correlate with a probable diagnosis, in ovarian cancer.
The markers of the present invention, alone or in combination, show a high degree of differential detection between ovarian cancer and non-cancerous states.
The markers of the present invention, alone or in combination, show a high degree of differential detection between ovarian cancer and non-cancerous states.
Description
DEMANDE OU BREVET VOLUMINEUX
LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND
PLUS D'UN TOME.
NOTE : Pour les tomes additionels, veuillez contacter le Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS
THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE THAN ONE
VOLUME
NOTE: For additional volumes, please contact the Canadian Patent Office NOM DU FICHIER / FILE NAME
NOTE POUR LE TOME / VOLUME NOTE:
DIFFERENTIAL EXPRESSION OF MARKERS IN OVARIAN CANCER
FIELD OF THE INVENTION
The present invention is related to novel nucleotide and protein sequences that are diagnostic markers for ovarian cancer, and assays and methods of use thereof.
BACKGROUND OF THE INVENTION
Ovarian cancer causes more deaths than any other cancer of the female reproductive system. An estimated 25,580 new cases will be diagnosed during 2004 in the United States, and approximately 16,090 of these women will die of the disease. Despite advances in the management of advanced ovarian cancer, 70% to 80% of patients will ultimately succumb to disease that is diagnosed in late stages. When ovarian cancer is diagnosed in stage I, more than 90% of patients can be cured with conventional surgery and chemotherapy. At present, however, only 25% of ovarian cancers are detected in stage I. Detection of a greater fraction lof ovarian l5 cancers at an early stage might significantly affect survival. A worldwide research effort, aiming at early detection of ovarian cancer, is currently being performed; finding molecular markers for the disease is one of the major research topics (J Clin Oncol. 2003 May 15;21(10 Suppl):200-5).
No single marker has been shown to be sufficiently sensitive or specific to contribute to the diagnosis of ovarian cancer. The marker that is currently most frequently used is CA-125 (Br J Cancer. 2000 May;82(9):1535-8). Its properties do not support its use for screening, but it is a major diagnostic tool. CA-125 is a member of the epithelial sialomucins markers group and is the most well documented and the best performing single marker from this group. Another name for CA-125 is mucin 16, and although it is a membrane protein, it can be found in the serum. Its greatest sensitivity is achieved for serous and emdometrioid ovarian tumors compared to mucinous or clear cell tumors. Other than diagnosis, it can be used for disease monitoring (Eur J
Gynaecol Oncol. 2000;21(1):64-9). In about 70% of patients, a rising level of CA-125 may be the first indication of relapse, predating clinical relapse by a median of 4 months. The serum concentration of CA-125 is elevated by the vascular invasion, tissue destruction and inflammation associated with malignant disease and is elevated in over 90% of those women with advanced ovarian cancer. Yet, CA-125 is not specific to ovarian cancer.
It is elevated in 40% of all patients with advanced intra-abdominal malignancy. Levels can also be elevated during menstruation or pregnancy and in other benign conditions such as cndometriosis, peritonitis or cirrhosis, particularly with ascites. CA-125 is not a marker that can be detected through use of urine samples due to a high molecular weight.
There are other ovarian cancer markers originating from epithelial mucins but none can replace CA-125, due to poorer specificity and sensitivity. These other markers may prove complementary to CA-125. CA-50, CA 54-61, CA-195 and CA 19-9 al l appear to have greater sensitivity for detection of mutinous tumors while STN and TAG-72 have better sensitivity for detection of clear cell tumors (Dis Markers. 2004;20(2):53-70).
Kallikreins, a family of serine proteases, and other protease-related proteins are also potential markers for ovarian cancer. Indeed, the entire family of kallikreins map to a region on chromosome 19q which is shown to be amplified in ovarian cancers. In particular, kallikrein 6 (protease M) and kallilrein 10 have been reported to have sensitivity up to 75% and specificity up to 100%. Matrix metalloproteinases (MMPs) are another family of proteases useful in ovarian cancer screening and prognosis. MMP-2 was reported to have 66%
sensitivity and 100%
specificity in one study. Cathepsin L, a cystein protease, was described to have a lower false positive rate compared with CA-125. Based on their biochemical proteolytic role, it would seem likely that these proteases would be active in invasion and metastasis formation and indeed these markers appear to have higher sensitivity for advanced stages of the disease.
Due to their relatively low molecular weight, such proteases are candidates to be urine markers, or markers which can be detected in urine samples (Dis Markers. 2004;20(2):53-70).
Hormones have a role in normal ovarian physiology. Therefore, it is not surprising that hormones, and growth and inhibition factors as well, are suitable for ovarian cancer detection.
Measurements of fragments of gonadotropin in the urine were found to have sensitivity up to 83% and specificity up to 92% for detecting ovarian cancer. Inhibins, members of the Transforming Growth Factors (TGF) beta superfamily, have been shown to have a diagnostic value in the detection of granulosa cell tumor, a relatively uncommon type of ovarian cancer, associated with better prognosis overall. Serum inhibin is an ovarian product which decreases to non detectable levels after menopause, however, certain ovarian cancers (mutinous carcinomas and sex cord stromal tumours such as granulosa cell tumours) continue to produce inhibin.
Studies have shown that that inhibin assays which detect all inhibin forms (as opposed to test detecting specific members of the inhibins family) provide the highest sensitivity/specificity characteristics as an ovarian cancer diagnostic test (Mol Cell Endocrinol.
2002 May 31; I 91 ( 1 ) :97-103). Measurement of serum TGF-alpha itself was found to have sensitivity up to 70% and specificity of 89% in early stage disease. The growth factor Mesothelin was also found to have diagnostic value but only for late stage disease.
Immunohistochemistry is frequently used to assess the origin of tumor and staging when a pathological tissue sample is available. A few molecular markers have been shown to have diagnostic value in Immunohistochemistry of ovarian cancer, among them Epidermal Growth Factor, p53 and HER-2. P53 expression is much lower at early stage than late stage disease. P53 high expression is more typical or characteristic of invasive serous tumors than of mucinous I 0 tumors. No benign tumors are stained with P53. HER-2 is found in less than 25% of newly diagnosed ovarian cancers. Ovarian cancer of type granulosa cell tumor has in general better prognosis with late relapse and/or metastasis formation. However, about 50% of patients still die within 20 years of diagnosis. In this specific tumor type, immunohistochemistry staining of estrogen receptor beta (ERb) and proliferating cell nuclear antigen (PCNA) showed that loss of ERb expression and high PCNA expression, characterized a subgroup of granulosa cell tumors with a worse outcome (Histopathology. 2003 Sep;43(3):254-62). Survivin expression was also shown to be correlated to tumor grade, histologic type and mutant p53 but actual correlation to survival is questionable (Mod Pathol. 2004 Feb;17(2):264) Many other markers have been tested over the years for ovarian cancer detection. Some markers have shown only limited value while others are still under investigation. Among them are TPA and TPS, two cytokeratins whose inclusion in a panel with CA-125 resulted in diagnoses with sensitivity up to 93% and specificity up to 98%. LPA -lysophosphatidic acid was a very promising marker with one study demonstrating 98% sensitivity and 90% specificity.
However, this marker is very unstable and requires quick processing and freezing of plasma, and 25. therefore has limited usage.
As previously described, no single marker has been shown to be sufficiently sensitive or specific to contribute to the diagnosis of ovarian cancer. Therefore combinations of markers in panel are being tested. Usually CA-125 is one of the panel members. The best performing panel combinations so far have been CA-125 with CA 15-3 with sensitivity of 93% and specificity of 93%, CA-125 with CEA (which has very little sensitivity by itself with specificity of 93% and specificity of 93%, and CA-125 with TAG-72 and CA 15-3 where specificity becomes 95% but sensitivity is diminished (Dis Markers. 2004;20(2):53-70).
SUMMARY OF THE INVENTION
The background art does not teach or suggest markers for ovarian cancer that are sufficiently sens hive and/or accurate, alone or in combination.
The present invention overcomes these deficiencies of the background art by providing novel markers for ovarian cancer that are both sensitive and accurate. These markers are differentially expressed and preferably overexpressed in ovarian cancer specifically, as opposed to normal ovarian tissue. The measurement of these markers, alone or in combination, in patient (biological) samples provides information that the diagnostician can correlate with a probable diagnosis of ovarian cancer. The markers of the present invention, alone or in combination, show a high degree of differential detection between ovarian cancer and norrcancerous states.
According to preferred embodiments of the present invention, examples of suitable biological samples which may optionally be used with preferred embodiments of the present invention include but are not limited to blood, serum, plasma, blood cells, urine, sputum, saliva, stool, spinal fluid or CSF, lymph fluid, the external secretions of the skin, respiratory, intestinal, and genitourinary tracts, tears, milk, neuronal tissue, ovarian tissue, any human organ or tissue, including any tumor or normal tissue, any sample obtained by lavage (for example of the bronchial system or of the female reproductive system), and also samples of in vivo cell culture constituents. In a preferred embodiment, the biological sample comprises ovarian tissue and/or a serum sample and/or a urine sample and/or secretions or other samples from the female reproductive system and/or any other tissue or liquid sample. The sample can optionally be diluted with a suitable eluant before contacting the sample to an antibody and/or performing any other diagnostic assay.
Information given in the text with regard to cellular localization was determined according to four different software programs: (i) tmhmm (from Center for Biological Sequence Analysis, Technical University of Denmark DTU, http://www.ebs.dtu.dk/services/TMHMM/TMHMM2.Ob.guide.php) or (ii) tmpred (from EMBnet, maintained by the ISREC Bionfonnatics group and the LICK Information Technology Office, Ludwig Institute for Cancer Research, Swiss Institute of Bioinfornatics, http://www.ch.embnet.org/software/TMPRED form.html) for transmembrane region prediction; (iii) signalp hmm or (iv) signalp nn (both from Center for Biological Sequence 5 Analysis, Technical University of Denmark DTU, http://www.cbs.dtu.dk/services/SignalP/background/prediction.php) for signal peptide prediction. The terms "signalp hmm" and "signalp nn" refer to two modes of operation for the program SignalP: hmm refers to Hidden Markov Model, while nn refers to neural networks.
Localization was also determined through manual inspection of known protein localization and/or gene structure, and the use of heuristics by the individual inventor.
In some cases for the manual inspection of cellular localization prediction inventors used the ProLoc computational platform [Einat Hazkani-Covo, Erez Levanon, Galit Rotman, Dan Graur and Amit Novik;
(2004) "Evolution of multicellularity in metazoa: comparative analysis of the subcellular localization of proteins in Saccharomyces, Drosophila and Caenorhabditis."
Cell Biology IS International 2004;28(3):171-8.], which predicts protein localization based on various parameters including, protein domains (e.g., prediction of trans-membranous regions and localization thereof within the protein), pI, protein length, amino acid composition, homology to pre-annotated proteins, recognition of sequence patterns which direct the protein to a certain organelle (such as, nuclear localization signal, NLS, mitochondria localization signal), signal peptide and anchor modeling and using unique domains from Pfam that are specific to a single compartment.
Information is given in the text with regard to SNPs (single nucleotide polymorphisms).
A description of the abbreviations is as follows. "T - > C", for example, means that the SNP
results in a change at the position given in the table from T to C. Similarly, "M - > Q", for example, means that the SNP has caused a change in the corresponding amino acid sequence, from methionine (M) to glutamine (Q). If, in place of a letter at the right hand side for the nucleotide sequence SNP, there is a space, it indicates that a frameshift has occurred. A
frameshift may also be indicated with a hyphen (-). A stop codon is indicated with an asterisk at the right hand side (*). As part of the description of an SNP, a comment may be found in parentheses after the above description of the SNP itself. This comment may include an FTId, which is an identifier to a SwissProt entry that was created with the indicated SNP. An FTId is a unique and stable feature identifier, which allows construction of links directly from position specific annotation in the feature table to specialized protein-related databases. The FTId is always the last component of a feature in the description field, as follows:
FTId=XXX number, in which XXX is the 3-letter code for the specific feature key, separated by an underscore from a Crdigit number. In the table of the amino acid mutations of the wild type proteins of the selected splice variants of the invention, the header of the first column is "SNP positions) on amino acid sequence", representing a position of a known mutation on amino acid sequence.
SNPs may optionally be used as diagnostic markers according to the present invention, alone or in combination with one or more other SNPs and/or any other diagnostic marker.
Preferred embodiments of the present invention comprise such SNPs, including but not limited to novel SNPs on the known (WT or wild type) protein sequences given below, as well as novel nucleic acid andlor amino acid sequences formed through such SNPs, and/or any SNP on a variant amino acid and/or nucleic acid sequence described herein.
Information given in the text with regard to the Homology to the known proteins was determined by SmitIrWaterman version 5.1.2 using special (non default) parameters as follows:
-model=sw.model -GAPEXT=0 -GAPOP=100.0 -MATRIX=blosuml 00 Information is given with regard to overexpression of a cluster in cancer based on ESTs.
A key to the p values with regard to the analysis of such overexpression is as follows:
- librar~based statistics: P-value without including the level of expression in cell-lines (P1) - library based statistics: P-value including the level of expression in cell-lines (P2) - EST clone statistics: P-value without including the level of expression in cell- lines (SP 1 ) - EST clone statistics: predicted overexpression ratio without including the level of expression in cell-lines (R3) - EST clone statistics: P-value including the level of expression in cell-lines (SP2) - EST clone statistics: predicted overexpression ratio including the level of expression in cell-lines (R4) Library based statistics refer to statistics over an entire library, while EST
clone statistics refer to expression only for ESTs from a particular tissue or cancer.
Information is given with regard to overexpression of a cluster in cancer based on microarrays. As a microarray reference, in the specific segment paragraphs, the unabbreviated tissue name was used as the reference to the type of chip for which expression was measured.
There are two types of microarray results: those from microarrays prepared according to a design by the present inventors, for which the microarray fabrication procedure is described in detail in Materials and Experimental Procedures section herein; and those results from microarrays using Affymetrix technology. As a microarray reference, in the specific segment paragraphs, the unabbreviated tissue name was used as the reference to the type of chip for which expression was measured. For microarrays prepared according to a design by the present I S inventors, the probe name begins with the name of the cluster (gene), followed by an identifying number. These probes are listed below with their respective sequences.
>H61775 0 11 0 CCCCAGCTTTTATAGAGCGGCCCAAGGAAGAATATTTCCAAGAAGTAGGG
>HSAPHOL 0 I1 0 GGAACATTCTGGATCTGACCCTCCCAGTCTCATCTCCTGACCCTCCCACT
>HUMGRPSE 0 0 16630 GCTGATATGGAAGTTGGGGAATCTGAATTGCCAGAGAATCTTGGGAAGAG
>HUMGRPSE 0 2 0 TCTCATAGAAGCAAAGGAGAACAGAAACCACCAGCCACCTCAACCCAAGG
>D56406 0 5 0 TCTGACTTTTACGGACTTGGCTTGTTAGAAGGCTGAAAGATGATGGCAGG
>M77904 0 8 0 AGTCTGTGTTTGAGGGTGAAGGCTCAGCAACCCTGATGTCTGCCAACTAC
>Z25299 0 3 0 AACTCTGGCACCTTGGGCTGTGGAAGGCTCTGGAAAGTCCTTCAAAGCTG
>Z44808 0 8 0 AAAAGCATGAGTTTCTGACCAGCGTTCTGGACGCGCTGTCCACGGACATG
>Z44808 0 0 72347 ATGTTCTTAGGAGGCAAGCCAGGAGAAGCCGGGTCTGACTTTTCAGCTCA
>Z44808 0 0 72349 TCCTCCAGACCCAAAGCCACAACCCATCGCAAGTCAAGAACACTTTCCAG
>S67314 0 0 741 CACAGAGCCAGGATGTTCTTCTGACCTCAGTATCTACTCCAGCTCCAGCT
>S67314 0 0 744 TGGCATGCTGGAACATGGACTCTAGCTAGCAAGAAGGGCTCAAGGAGGTG
>Z39337 0 0 66755 GCAGGGGTTAAAAGGACGTTCCAGAAGCATCTGGGGACAGAACCAGCCTC
>Z39337 0 9 0 TAATAAACGCAGCGACGTGAGGGTCCTGATTCTCCCTGGTTTTACCCCAG
>HUMPHOSLIP 0 0 18458 AAGGAAGCAGGACCAGTGGATGTGAGGCGTGGTCGAAGAACAACAGAAAG
>HUMPHOSLIP 0 0 18487 ACAGGGGCCAGATGGTGACCCATGACCCAGCCTAAAAGGCAGCCAGAGGG
>M78530 0 6 0 CTTCCTACACACATCTAGACGTTCAAGTTTGCAAATCAGTTTTTAGCAAG
>HSMUC1A 0 37 0 AAAAGGAGACTTCGGCTACCCAGAGAAGTTCAGTGCCCAGCTCTACTGAG
>HSMUC1A 0 0 11364 AAAGGCTGGCATAGGGGGAGGTTTCCCAGGTAGAAGAAGAAGTGTCAGCA
>HSMUC 1 A 0 0 11365 AATTAACCCTTTGAGAGCTGGCCAGGACTCTGGACTGATTACCCCAGCCT
Oligonucleotide microarray results taken from Affymetrix data were from chips available from Affymetrix Inc, Santa Clara, CA, USA (see for example data regarding the Human Genome U133 (HG-U133) Set at www.affymetrix.com/products/arrays/specific/hgul33.affx; GeneChip Human Genome 2.0 Array at www.affymetrix.com/products/arrays/specific/hgu133av2.affx; and Human Genome U133 Plus 2.0 Array at www.affymetrix.com/products/arrays/specific/hgu I 33plus.affx). The probe names follow the Affymetrix naming convention. The data is available from NCBI Gene Expression Omnibus (see www.ncbi.nlm.nih.gov/projects/geo/ and Edgar et al, Nucleic Acids Research, 2002, Vol.
30, No. 1 207-210). The dataset (including results) is available from www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE 1133 for the Series GSE 1133 database (published on March 2004); a reference to these results is as follows: Su et al (Proc Natl Acad Sci U S A. 2004 Apr 20;1 O 1 ( I 6):6062-7. Epub 2004 Apr 09).
The following list of abbreviations for tissues was used in the TAA
histograms. The term l0 "TAA" stands for "Tumor Associated Antigen", and the TAA histograms, given in the text, represent the cancerous tissue expression pattern as predicted by the biomarkers selection engine, as described in detail in examples 1-5 below (the first word is the abbreviation while the second word is the full name):
("BONE", "bone");
("COL", "colon");
("EPI", "epithelial");
("GEN", "general");
("LIVER", "liver");
(~~L~~~~ ~~lung~~)~
("LYMPH", "lymph nodes");
("MARROW", "bone marrow");
("OVA", "ovary");
("PANCREAS", "pancreas");
("PRO", "prostate");
("STOMACH", "stomach");
("TCELL", "T cells");
("THYROID", "Thyroid");
("MAM", "breast");
(..BRAIN.,, ..brain");
("UTERUS", "uterus");
("SKIN", "skin");
("KIDNEY", "kidney");
("MUSCLE", "muscle");
("ADREN", "adrenal");
("HEAD", "head and neck");
5 ("BLADDER", "bladder");
It should be noted that the terms "segment", "seg" and "node" are used interchangeably in reference to nucleic acid sequences of the present invention; they refer to portions of nucleic 10 acid sequences that were shown to have one or more properties as described below. They are also the building blocks that were used to construct complete nucleic acid sequences as described in greater detail below. Optionally and preferably, they are examples of oligonucleotides which are embodiments of the present invention, for example as amplicons, hybridization units and/or from which primers and/or complementary oligonucleotides may optionally be derived, and/or for any other use.
Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this invention belongs. The following references provide one of skill with a general definition of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed.
1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988);
The Glossary of Genetics, Sth Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). All of these are hereby incorporated by reference as if fully set forth herein. As used herein, the following terms have the meanings ascribed to them unless specified otherwise.
As used herein the phrase "ovarian cancer" refers to cancers of the ovary including but not limited to Ovarian epithelial tumors (serous, mucinous, endometroid, clear cell, and Brenner tumor), ovarian germ-cell tumors, (teratoma, dysgerminoma, endodermal sinus tumor, and embryonal carcinoma) and ovarian stromal tumors (originating from granulosa, theca, Sertoli, Leydig, and collagerrproducing stromal cells).
m The teen "marker" in the context of the present invention refers to a nucleic acid fragment, a peptide, or a polypeptide, which is differentially present in a sample taken from subjects (patients) having ovarian cancer as compared to a comparable sample taken from subjects who do not have ovarian cancer.
The phrase "differentially present" refers to differences in the quantity of a marker present in a sample taken from patients having ovarian cancer as compared to a comparable sample taken from patients who do not have ovarian cancer. For example, a nucleic acid fragment may optionally be differentially present between the two samples if the amount of the nucleic acid fragment in one sample is significantly different from the amount of the nucleic acid fragment in the other sample, for example as measured by hybridization andlor NAT-based assays. A polypeptide is differentially present between the two samples if the amount of the polypeptide in one sample is significantly different from the amount of the polypeptide in the other sample. It should be noted that if the marker is detectable in one sample and not detectable in the other, then such a marker can be considered to be differentially present.
As used herein the phrase "diagnostic" means identifying the presence or nature of a pathologic condition. Diagnostic methods differ in their sensitivity and specificity. The "sensitivity" of a diagnostic assay is the percentage of diseased individuals who test positive (percent of "true positives"). Diseased individuals not detected by the assay are "false negatives." Subjects who are not diseased and who test negative in the assay are termed "true negatives." The "specificity" of a diagnostic assay is I minus the false positive rate, where the "false positive" rate is defined as the proportion of those without the disease who test positive.
While a particular diagnostic method may not provide a definitive diagnosis of a condition, it suffices if the method provides a positive indication that aids in diagnosis.
As used herein the phrase "diagnosing" refers to classifying a disease or a symptom, determining a severity of the disease, monitoxing disease progression, forecasting an outcome of a disease and/or prospects of recovery. The term "detecting" may also optionally encompass any of the above.
Diagnosis of a disease according to the present invention can be effected by determining a level of a polynucleotide or a polypeptide of the present invention in a biological sample obtained from the subject, wherein the level determined can be correlated with predisposition to, or presence or absence of the disease. It should be noted that a "biological sample obtained from the subject" may also optionally comprise a sample that has not been physically removed from the subject, as described in greater detail below.
As used herein, the term "level" refers to expression levels of RNA and/or protein or to DNA copy number of a marker of the present invention.
Typically the level of the marker in a biological sample obtained from the subject is different (i.e., increased or decreased) from the level of the same variant in a similar sample obtained from a healthy individual (examples of biological samples are described herein).
Numerous well known tissue or fluid collection methods can be utilized to collect the biological sample from the subject in order to determine the level of DNA, RNA
and/or polypeptide of the variant of interest in the subject.
Examples include, but are not limited to; fine needle biopsy, needle biopsy, core needle biopsy and surgical biopsy (e.g., brain biopsy), and lavage. Regardless of the procedure employed, once a biopsy/sample is obtained the level of the variant can be determined and a diagnosis can thus be made.
Determining the level of the same variant in normal tissues of the same origin is preferably effected along-side to detect an elevated expression and/or amplification and/or a decreased expression, of the variant as opposed to the normal tissues.
A "test amount" of a marker refers to an amount of a marker in a subject's sample that is consistent with a diagnosis of ovarian cancer. A test amount can be either in absolute amount (e.g., microgram/ml) or a relative amount (e.g., relative intensity of signals).
A "control amount" of a marker can be any amount or a range of amounts to be compared against a test amount of a marker. For example, a control amount of a marker can be the amount of a marker in a patient with ovarian cancer or a person without ovarian cancer. A
control amount can be either in absolute amount (e.g., microgram/ml) or a relative amount (e.g., relative intensity of signals).
"Detect" refers to identifying the presence, absence or amount of the object to be detected.
A "label" includes any moiety or item detectable by spectroscopic, photo chemical, biochemical, immunochemical, or chemical means. For example, useful labels include 32p, 3sS, fluorescent dyes, electrorrdense reagents, enzymes (e.g., as commonly used in an ELISA), biotin-streptavadin, dioxigenin, haptens and proteins for which antisera or monoclonal antibodies are available, or nucleic acid molecules with a sequence complementary to a target.
The label often generates a measurable signal, such as a radioactive, chromogenic, or fluorescent signal, that can be used to quantify the amount of bound label in a sample. The label can be incorporated in or attached to a primer or probe either covalently, or through ionic, van der Waals or hydrogen bonds, e.g., incorporation of radioactive nucleotides, or biotinylated nucleotides that are recognized by streptavadin. The label may be directly or indirectly detectable. Indirect detection can involve the binding of a second label to the first label, directly or indirectly. For example, the label can be the ligand of a binding partner, such as biotin, which is a binding partner for streptavadin, or a nucleotide sequence, which is the binding partner for a complementary sequence, to which it can specifically hybridize. The binding partner may itself be directly detectable, for example, an antibody may be itself labeled with a fluorescent molecule. The binding partner also may be indirectly detectable, for example, a nucleic acid having a complementary nucleotide sequence can be a part of a branched DNA
molecule that is in turn detectable through hybridization with other labeled nucleic acid molecules (see, e.g., P.
D. Fahrlander and A. Klausner, Bio/Technology 6:1165 (1988)). Quantitation of the signal is achieved by, e.g., scintillation counting, densitometry, or flow cytometry.
Exemplary detectable labels, optionally and preferably for use with immunoassays, include but are not limited to magnetic beads, fluorescent dyes, radiolabels, enzymes (e.g., horse radish peroxide, alkaline phosphatase and others commonly used in an ELISA), and calorimetric labels such as colloidal gold or colored glass or plastic beads.
Alternatively, the marker in the sample can be detected using an indirect assay, wherein, for example, a second, labeled antibody is used to detect bound marker-specific antibody, and/or in a competition or inhibition assay wherein, for example, a monoclonal antibody which binds to a distinct epitope of the marker are incubated simultaneously with the mixture.
"Immunoassay" is an assay that uses an antibody to specifically bind an antigen. The immunoassay is characterized by the use of specific binding properties of a particular antibody to isolate, target, andlor quantify the antigen.
The phrase "specifically (or selectively) binds" to an antibody or "specifically (or selectively) immunoreactive with," when referring to a protein or peptide (or other epitope), refers to a binding reaction that is determinative of the presence of the protein in a heterogeneous population of proteins and other biologics. Thus, under designated immunoassay conditions, the specified antibodies bind to a particular protein at least two times greater than the background (non-specific signal) and do not substantially bind in a significant amount to other proteins present in the sample. Specific binding to an antibody under such conditions may require an antibody that is selected for its specificity for a particular protein. For example, polyclonal antibodies raised to seminal basic protein from specific species such as rat, mouse, or human can be selected to obtain only those polyclonal antibodies that are specifically immunoreactive with seminal basic protein and not with other proteins, except for polymorphic variants and alleles of seminal basic protein. This selection may be achieved by subtracting out antibodies that cross-react with seminal basic protein molecules from other species. A variety of immunoassay formats may be used to select antibodies specifically immunoreactive with a particular protein. For example, solid-phase ELISA immunoassays are routinely used to select antibodies specifically immunoreactive with a protein (see, e.g., Harlow &
Lane, Antibodies, A
Laboratory Manual (1988), for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity). Typically a specific or selective reaction will be at least twice background signal or noise and more typically more than 10 to 100 times background.
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~,...
~T'ranscript Name ;u~ a.. ,. ..
z H571775 T21 .
a nucleic acid sequence comprising a sequence in the table below:
'ur .~ ~ ' a,.
>,'~Segme°n~C Name -~ ~ , ø._._. x H617?5 node 2 H61775 node 4 H61775 node 6 H61775 node 8 H61775 node 0 H61775 node 5 According to preferred embodiments of the present invent ion, there is provided an isolated polypeptide comprising an amino acid sequence in the table below amino acid sequence comprising a sequence in the table below:
Prot~ln~Naane ~:'~ ~~'~
- ~ -~
~ : ~~ v ~. ..~, ~z ..s,::%~ :m."~..,.~-.'-~;
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
Transcript~ame - ~' ~
~
~~, ~ ~~- ~_.
L ~~.~
10 a nucleic acid sequence comprising a sequence in the table below:
Se ent Name ~
~$
~' ''~~s ~~~
HUMCEA PEA 1 node 0 HUMCEA PEA 1 node 2 HUMCEA PEA 1 node 11 HUMCEA PEA 1 node 12 HUMCEA PEA 1 node 31 HUMCEA PEA I
node node node node node node node node node HUMCEA PEA node9 I
HUMCEA PEA node10 HUMCEA PEA node15 HUMCEA PEA node16 HUMCEA PEA node17 I
HUMCEA PEA node18 HUMCEA PEA node19 HUMCEA PEA node20 HUMCEA PEA node21 I
HUMCEA PEA node22 HUMCEA PEA node23 HUMCEA PEA node24 HUMCEA PEA node27 HUMCEA PEA node29 HUMCEA PEA node30 HUMCEA PEA node33 HUMCEA PEA node34 HUMCEA PEA node35 HUMCEA PEA node HUMCEA PEA node50 HUMCEA PEA node51 HUMCEA PEA node 56 HUMCEA PEA node 57 I
HUMCEA PEA node 58 HUMCEA PEA node 60 HUMCEA PEA node 61 HUMCEA PEA node 62 HUMCEA PEA-1node 64 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
w,-.~ . ~,.
~.>.. ,.~ w.
~ . ~.. ,Y,.:.~ . ~~ -.~., (~'~~ x ~-..v~,.. ~~orres ondm . Traris~n ~ s s, :
.- .
Protem~Name~ ~x , _, .., ~ ~- ..~
~_~ C~' .:- 9 ' , v. ~
s;~~~ , ' ~
'' .a3:..,.,<. ~
-..a , "
-~,~ , v ~ ..," ~
y~ W"-v~ ,.
~" a:''.~"a=;: ....
: ....
~ sY,,.-x~-"
x,;.
t ~r~ a.~ ": .,~...,~'~:.;-_:~
>. rc.
~
~~
- > ... ':. ~ .,~..~:~".:.,.",.._ ~ ~...~,<> x. _,>a"5-~,. ~.:~-~"~;;;
M
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~'zan~crt'~1~1~~'rne~~~~~~~.
s _ ~
~'~
~
~
'~ ..:..
, .
:
~
a nucleic acid sequence comprising a sequence in the table below:
~Segnient~Narine' '' "'_'~b ,. ~"-HUMEDF PEA 2node6 1-IUMEDF 2node11 PEA
HUMEDF PEA 2nodet 8 H UMEDF 2node19 PEA
HUMEDF PEA 2node22 I-IUMEDF 2node2 PEA
HUMEDF PEA 2node8 I-IUMEDF 2node20 PEA
According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
.,.;e;?';',.~~~' a; ,.:;... ",~'~~.a<-..~.. . , . ~" w~~
..~r,w ...,., a.. .~3""..- .~.~.;....~~'-, ,~a7~ '.." s. .. a. "~ '"~ ,.,~.r~
~L'rotena Name~~~~= -.:~ ~..- , ." -: ~-..r:..f.:.
~.-3 ~ CoiTes on_d~n - ~z~nscr~ t a ~.~v ~;..,: ,;~ .- ":.... :,>. .... ~ ~ o.~ ~..:.,"".
: t ~: ~R
~ a, ~- ~
~? ~
, MY a h 4 ' :~
~W
~
-.
Y
i .."",...; ,..
" ~.
,.. ,. ,~.C
~~m.,.. < ",.w, W~a.
, : s, ..., ~
~,. ~....,.
.,. .., .
,.,.. 7 ..,..
.,.
.
, , ..
.
.
r ~
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below andlor:
s~~': ~ ~. ~r ~~ ~'~~Cl 1~~a?T~~ a~ ~
~.- :,- ~- ~~ , a~
HSAPHOL T10 ~~
HSAPHOL TS
a nucleic acid sequence comprising a sequence in the table below:
~ ~-s ~a~--,_ ~ ~s:.:.' .n.-...-,~. x~ .s m -;-:
Segment Name.
-''~
HSAPHOL node 1 l Y
I-ISAPHOL node 13 HSAPHOL node 15 HSAPHOL node 19 HSAPHOL node 2 HSAPI-IOL node 21 HSAPHOL node 23 HSAPHOL node 26 HSAPHOL node 28 HSAPHOL node 38 HSAPHOL node 40 HSAPHOL node 42 HSAPHOL node 16 HSAPHOL node 25 HSAPHOL node 34 HSAPHOL node 35 HSAPHOL node 36 HSAPHOL node 41 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
r ~1 a~e,w 3 ~~
2o HSAPHOL PS
~HSAPHOL P8 According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
,.m ~ ~ ~ ~--_ °~ f Traris rlpt Name ~ .
y, z ~ ~ w~ . a ;' .'~:._.. r . ~ 2.~.,..?. ~ :es a nucleic acid sequence comprising a sequence in the table below:
Sgt~~en~lame m b ~ y T1088 I ll PEA node node node node node node node node-According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
a ~aa a '~,~ t ~.Y~' ':~ ~t s,~' ~ ~ ~s "' ~P~rotem Name~~~~ ~ F~~ ~~~ ~~Y, , ~.
~. x,7 - X-~..,v ..,~-..»~ .~°.~,.,r... <s~'.e. '~.- , . ,a ~
.~ :_ss T10888 PEA_1 P6 According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~~~ ~r .~
~ ~ ~r scr~pt ~ .~ ~w Nam e ~
~ ~ ~
_ .".R~. ~..a ~,.~.-,~
~ ~ _z ~,. ,.a '.~ . ",,m",~a.
~ m~ , .~,s l a nucleic acid sequence comprising a sequence in the table below:
~~S~e ~i~~it am ~ ~~~~ -'_' ~ ~ ~ ~ x ~,.t~a ~3',.X~a g a F, ~" i~~ na,c ~. A7,'~.
~ z~, . . ? " ~4.. , F~,r,.,B~'I~.lhf,r HSECADH node 0 I-ISECADH node 14 HSECADH node 15 HSECADH node 21 HSECADH node 22 HSECADH node 25 HSECADH node 26 HSECADH node 48 HSECADH node 52 HSECADH node 53 HSECADH node 54 HSECADH node 57 HSECADH node 60 HSECADH node 62 HSECADH node 63 HSECADH node 7 HSECADH node 1 HSECADH node 11 HSECADH node 12 HSECADH node 17 HSECADH node 18 HSECADH node 19 HSECADH node 3 HSECADH node HSECADH node HSECADH node HSECADH node 55 HSECADH node 56 HSECADH node HSECADH node 59 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~~ iltein~Nam~
., a -_s ~ a": a'~.R :. . zoo .
HSECADH P l 5 According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~" ~
Transcript Name- ~--~-~' ~-~ ~ ~-y HUMGRPSE TS
a nucleic acid sequence comprising a sequence in the table below:
~~ Se~gm~e~'nt~~ame a ~~
W
E
rv HUMGRPSE node 0 HUMGRPSE node .
HUMGRPSE node 8 HUMGRPSE node 3 HUMGRPSE node 7 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
Protern~Name~ _ ~ ~ -~ ,~~ ~~
HUMGRPSE PS
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
y Transcri t Na ne p l ~.. ~ _.
R 11723 I T l 7 PEA
PEA
a nucleic acid sequence comprising a sequence in the table below:
~'~.Segrnent~I~am~~~
~~_ p. ~
~_ ~:. .
..~:~, ~F~o._ T: .-.~
-. ., _ W~
811723 PEA l 13 node node node node node node node node node node node node node node node node node 811723 PEA_1 node 811723 PEA node29 R 1 1723PEA node3 R 1 1723PEA node30 R 11723 PEA node4 811723 PEA node5 811723 PEA node6 811723 PEA node7 I
811723 PEA node8 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~,Protem~N~xn~ '~'~ ~~ ~~"xr ~ R
~~:: ~:~,a ,,~_..,._.....:~»..
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~ ~ ~ ~~ . x~, ~~anscri t~~Tama ~~ ~ri ~''~" ~w~.~=g ~ ~ ~,~~ .~
_ ~.~ _~
D56406 PEA 1 T3 ~
a nucleic acid sequence comprising a sequence in the table below:
a, .. . - ...,,.
, a Segment Nalne ,:
a;
~-.
v '..~
a"~~
D56406 PEA node0 I
D56406 PEA node13 D56406 PEA node11 D56406 PEA node2 D56406 PEA node3 D56406 PEA node5 D56406 PEA node6 I
D56406 PEA node7 DS6406 PEA node8 I
D56406_PEA_1 node9 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
rCf>rf31n1'~t~llle~ '~-~~ _ :~, a .,~ r<
According to preferred embodiments ofthe present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
T ~scz~p Name ~ ~ ~ , ~, _, H53393 PEA~1 T9 a nucleic acid sequence comprising a sequence in the table below:
,_ ~ .Segment Name ~ ~
s ~...m~
. E.~
axe~~.>.~
~ ~
x -'.
v ~e H53393 PEA node 0 I
H53393 PEA node 10 H53393 PEA node 12 H53393 PEA node 13 I
H53393 PEA node 1 S
I
H53393 PEA node 17 H53393 PEA node 19 I
H53393 PEA node 23 I
H53393 PEA node 24 H53393 PEA node 25 H53393 PEA node 29 I
H53393 PEA node 4 H53393 PEA node 6 H53393 PEA node 8 I
H53393 PEA node 21 H53393 PEA node 22 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
" .. .., ; -:~ w~~;'m'.o.,. ~1' . .. _~~~
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below andlor:
~~TraiascriptNat~e~' '~ ~ ~ v~~~~.
;~ ~ . ~- ; ~ _ -~-a a _ ~. _ .. , -~ ~f _ ,~-.~ ~.
a nucleic acid sequence comprising a sequence in the table below:
~~Segment~Name~-~
~ ..~~~~
-~~ '--.~' ,.
~
HSU40434 PEA Inode l HSU40434 PEA 1node 16 HSU40434 PEA Inode 30 HSU40434 PEA Inode 32 HSU40434 PEA Inode 57 HSU40434 PEA Inode 0 HSU40434 PEA 1node 10 HSU40434 PEA 1node 13 HSU40434 PEA 1node 18 HSU40434 PEA 1node 2 HSU40434 PEA 1node 20 HSU40434 PEA 1node 21 HSU40434 PEA 1node 23 HSU40434 PEA 1node 24 HSU40434 PEA 1node 26 HSU40434 PEA 1node 28 HSU40434 PEA 1node 3 HSU40434 PEA 1node 35 HSU40434 PEA 1node 36 HSU40434 PEA 1node 37 HSU40434 PEA 1node 38 I-ISU40434 PEA node 39 I
HSU40434 PEA node 40 HSU40434 PEA node 41 HSU40434 PEA node 42 HSU40434 PEA node 43 I
HSU40434 PEA node 44 HSU40434 PEA node 47 I
HSU40434 PEA node 48 HSU40434 PEA node 51 HSU40434 PEA node 52 HSU40434 PEA node 53 HSU40434 PEA node 54 I
HSU40434 PEA node 56 HSU40434 PEA node 7 HSU40434 PEA-1 node-8 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
PrOt~81r1~ ~ ~ '~~~;~~~ ~- " S
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~' 'I~rans ~t TT~~ ne ~z x, .~~,~ ~ _:
a nucleic acid sequence comprising a sequence in the table below:
~_ ~~ ~ gment~larne~ ~ ~ ~ ~~ ~ ~ ~ ~=, ~, ~ø_ -~ w....~..__.,.
M77904 node 0 M77904 node 11 M77904 node 12 M77904 node 14 M77904 node 15 M77904 node 17 M77904 node 2 M77904 node 21 M77904 node 23 M77904 node 24 M77904 node 27 M77904 node 28 M77904 node 4 M77904 node 6 M77904 node 7 M77904 node 8 M77904 node 9 M77904 node 19 M77904 node 22 M77904 node 25 M77904 node 26 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
Protein e ~~ ' ~ : ~ ~ '~m~.
Nam ~~ ~~
s ~ a ..~
P , -; "~: _ -~i=Tv'- ;~--.
'',n_aa;,. , '," e' -...
PS
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~, ~
'Txanscnpt Name ~_ ~ ~ >~~ ~ ~ ri~
_,.,. ~, ~ "~ r,p.
s' .. ;':~ ~":, .«.., Wit' 's '~" , ~ ''~;_,' .~,a g a nucleic acid sequence comprising a sequence in the table below:
~egment~
am ~ x T
~~ ~ ~ "
~m.
w _..~ r w_~,.;~
'~z :'8~
:~ ~ 'a~s.~:y'~i~~
~,'~ . ,.
~ ~ .z:"
225299 PEA 2 node 20 225299 PEA 2 node 21 225299 PEA 2 node 23 225299 PEA 2 node 24 225299 PEA 2 node 8 225299 PEA 2 node 12 2 node 13 225299 PEA 2 node 14 2 node 17 225299 PEA 2 node 18 225299 PEA 2 node 19 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
Pr_oteinNa~ie ~~ ~~.~~~~ ~~_~r ~, ' g~ ~, ='~
Z25299~ PEA 2 P2 According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
TIaIi~Cl'l~t NalTte~° '~x~' ~ a, T39971 T1"0 a nucleic acid sequence comprising a sequence in the table below:
a en ame ~~
~'~..~' ~
' ~
,~
.;..
,:
T39971node0 T39971node18 T39971node21 T39971node22 T39971node23 T39971node31 T39971node33 9971 node7 T39971node1 T39971node10 T39971node11 T39971node12 T39971node15 T39971node16 T39971node17 T39971node26 T39971node27 T39971node28 T39971node29 T39971node3 T39971node30 T39971node34 T39971node35 T39971node36 T39971node4 T39971node5 T39971node8 T39971node9 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
Protein Name ~ ~~~i ~'~~ °" °~ ~ ~ ~:
f A~~~~~.,n.~ o According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~ a s...
Transcript Name~~~- ~ ~~m ~~ ~.
244808 .PEA 1 T5 a nucleic acid sequence comprising a sequence in the table below:
~Se~grnent~Name 244808PEAI node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808 1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node II
244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA 1 node 35 244808PEA 1 node 39 244808PEA node 4 I
244808PEA node 6 44808PEA-1node 8 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
Paz ~P~,rote~n Nar~n'e ~,~
_s ~
~k,..;;...~~,~ . r,.~.''.~_ . ~....,~... .~" ~ ~y,,r '~:, According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~TrdIISCrI~t ~8 "' "s~~"w a nucleic acid sequence comprising a sequence in the table below:
Segra~ent Name~~
' ~
-P' T
g -:
.~k~
.
bh~.,...<.~"K9 ~,.'~.~,'.~5r ' S67314 node 0 PEA
567314PEA node 11 I
567314PEA node 13 567314PEA node 15 l 567314PEA 1 node 17 567314PEA node 4 S67314PEA node 10 567314PEA node 3 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
otem Name ~ ~
~ ~' ~~ ~ z ~:
' ' ~
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~'P a scnpt~ a"nie~°~ '~~"~~c ~x~ ~~ ~:u ~3 ~~
.~ _~~.-.~ .
239337 PEA t2 PEA 1 T3 239337 PEA 2 PEA_1 T12 a nucleic acid sequence comprising a sequence in the table below:
~~t N~_ tee . ;
~ ~ y ~ a 2 .. ,~T.,'~
_~..~'' ' ~,.i., 239337 PEA PEA node 2 239337 PEA PEA_1node-15 239337 PEA PEA_1node_16 239337 PEA PEA node 18 239337 PEA PEA node 21 239337 PEA PEA-1node 22 239337PEA 2 PEA node 3 239337PEA 2 PEA node 5 239337PEA 2 PEA node 6 239337PEA 2 PEA node 10 239337PEA 2 PEA node 11 239337 2 PEA node 14 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
z,:.xa. .~ ,:; .x ' ~.~, ~ 4'~~:...
~ .~,;;-:3~.~ s .~....'~~ ,x. -:-~ ., sa .
vs , rs ,.::::.-~',";cx;r ,<;,.,a" n ra , ,~;~.,:~t,,,."..w'~
s ,.3",??
r ."'..s:' ., .,?.':'. "~ .l:~c: ~ tea- ~ a ae. ' ,., a ;~ : .,1 Prot~mName~~~.~ ~~. _ _~w_ w~ ~ r ~~ ~~ ,~ ~_~~~.-~ ~' . ~, ~
a ~~
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
Txanscnp aye ..,..~.':.~-o.~.
,~~'f~' s~
~3 ' M Y~
~ E~ i~ ~Hf ~~
PEA
PEA
PEA
PEA
PEA
a nucleic acid sequence comprising a sequence in the table below:
~5entN~'~aine ~~, ~- ~~ ~ ~ ,-u-~:
~w ~..:. ~. _ ~~.,~ .; a HUMPHOSLIP PEA 2 node 0 HUMPHOSLIP 2 node19 PEA
HUMPHOSLIP 2 node34 PEA
HUMPHOSLIP 2 node68 PEA
HUMPHOSLIP 2 node70 PEA
HUMPHOSL1P 2 node75 PEA
HUMPHOSLIP 2 node2 PEA
HUMPHOSLIP 2 node3 PEA
HUMPHOSLIP 2 node HUMPHOSLIP 2 node6 PEA
HUMPHOSLIP 2 node7 PEA
HUMPHOSLIP 2 node8 PEA
HUMPHOSLIP 2 node9 PEA
HUMPHOSLIP 2 nodeI4 PEA
HUMPHOSLIP 2 node15 PEA
HUMPHOSLIP 2 node16 PEA
HUMPHOSLIP 2 node17 PEA
HUMPHOSLIP node23 HUMPHOSLIP node24 HUMPHOSLIP 2 node HUMPHOSLIP 2 node26 PEA
HUMPHOSLIP 2 node PEA node HUMPHOSLIP node33 HUMPHOSLIP node36 HUMPHOSLIP node37 HUMPHOSLIP node39 HUMPHOSLIP node HUMPHOSLIP node HUMPHOSLIP node HUMPHOSLIP 2 node HUMPI-IOSLIP node 45 HUMPHOSLIP PEA node 47 HUMPHOSLIP PEA node 51 HUMPHOSLIP PEA node 52 HUMPHOSLIP PEA node 53 HUMPHOSLIP PEA node 54 HUMPHOSLIP PEA node 55 HUMPHOSLIP PEA node 58 HUMPHOSLIP PEA node 59 HUMPHOSLIP PEA node 60 HUMPHOSLIP PEA node 61 HUMPHOSLIP PEA node 62 HUMPHOSLIP PEA node 63 HUMPHOSLIP PEA node 64 HUMPHOSLIP PEA node 65 HUMPHOSLIP PEA node 66 HUMPHOSLIP PEA node 67 HUMPHOSLIP PEA node 69 HUMPHOSLIP PEA node 71 HUMPHOSLIP PEA node 72 .2 HUMPHOSLIP PEA node 73 HUMPHOSLIP PEA node 74 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
PEA
PEA
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~Tr nsc pt Named ~
a nucleic acid sequence comprising a sequence in the table below:
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DIFFERENTIAL EXPRESSION OF MARKERS IN OVARIAN CANCER
FIELD OF THE INVENTION
The present invention is related to novel nucleotide and protein sequences that are diagnostic markers for ovarian cancer, and assays and methods of use thereof.
BACKGROUND OF THE INVENTION
Ovarian cancer causes more deaths than any other cancer of the female reproductive system. An estimated 25,580 new cases will be diagnosed during 2004 in the United States, and approximately 16,090 of these women will die of the disease. Despite advances in the management of advanced ovarian cancer, 70% to 80% of patients will ultimately succumb to disease that is diagnosed in late stages. When ovarian cancer is diagnosed in stage I, more than 90% of patients can be cured with conventional surgery and chemotherapy. At present, however, only 25% of ovarian cancers are detected in stage I. Detection of a greater fraction lof ovarian l5 cancers at an early stage might significantly affect survival. A worldwide research effort, aiming at early detection of ovarian cancer, is currently being performed; finding molecular markers for the disease is one of the major research topics (J Clin Oncol. 2003 May 15;21(10 Suppl):200-5).
No single marker has been shown to be sufficiently sensitive or specific to contribute to the diagnosis of ovarian cancer. The marker that is currently most frequently used is CA-125 (Br J Cancer. 2000 May;82(9):1535-8). Its properties do not support its use for screening, but it is a major diagnostic tool. CA-125 is a member of the epithelial sialomucins markers group and is the most well documented and the best performing single marker from this group. Another name for CA-125 is mucin 16, and although it is a membrane protein, it can be found in the serum. Its greatest sensitivity is achieved for serous and emdometrioid ovarian tumors compared to mucinous or clear cell tumors. Other than diagnosis, it can be used for disease monitoring (Eur J
Gynaecol Oncol. 2000;21(1):64-9). In about 70% of patients, a rising level of CA-125 may be the first indication of relapse, predating clinical relapse by a median of 4 months. The serum concentration of CA-125 is elevated by the vascular invasion, tissue destruction and inflammation associated with malignant disease and is elevated in over 90% of those women with advanced ovarian cancer. Yet, CA-125 is not specific to ovarian cancer.
It is elevated in 40% of all patients with advanced intra-abdominal malignancy. Levels can also be elevated during menstruation or pregnancy and in other benign conditions such as cndometriosis, peritonitis or cirrhosis, particularly with ascites. CA-125 is not a marker that can be detected through use of urine samples due to a high molecular weight.
There are other ovarian cancer markers originating from epithelial mucins but none can replace CA-125, due to poorer specificity and sensitivity. These other markers may prove complementary to CA-125. CA-50, CA 54-61, CA-195 and CA 19-9 al l appear to have greater sensitivity for detection of mutinous tumors while STN and TAG-72 have better sensitivity for detection of clear cell tumors (Dis Markers. 2004;20(2):53-70).
Kallikreins, a family of serine proteases, and other protease-related proteins are also potential markers for ovarian cancer. Indeed, the entire family of kallikreins map to a region on chromosome 19q which is shown to be amplified in ovarian cancers. In particular, kallikrein 6 (protease M) and kallilrein 10 have been reported to have sensitivity up to 75% and specificity up to 100%. Matrix metalloproteinases (MMPs) are another family of proteases useful in ovarian cancer screening and prognosis. MMP-2 was reported to have 66%
sensitivity and 100%
specificity in one study. Cathepsin L, a cystein protease, was described to have a lower false positive rate compared with CA-125. Based on their biochemical proteolytic role, it would seem likely that these proteases would be active in invasion and metastasis formation and indeed these markers appear to have higher sensitivity for advanced stages of the disease.
Due to their relatively low molecular weight, such proteases are candidates to be urine markers, or markers which can be detected in urine samples (Dis Markers. 2004;20(2):53-70).
Hormones have a role in normal ovarian physiology. Therefore, it is not surprising that hormones, and growth and inhibition factors as well, are suitable for ovarian cancer detection.
Measurements of fragments of gonadotropin in the urine were found to have sensitivity up to 83% and specificity up to 92% for detecting ovarian cancer. Inhibins, members of the Transforming Growth Factors (TGF) beta superfamily, have been shown to have a diagnostic value in the detection of granulosa cell tumor, a relatively uncommon type of ovarian cancer, associated with better prognosis overall. Serum inhibin is an ovarian product which decreases to non detectable levels after menopause, however, certain ovarian cancers (mutinous carcinomas and sex cord stromal tumours such as granulosa cell tumours) continue to produce inhibin.
Studies have shown that that inhibin assays which detect all inhibin forms (as opposed to test detecting specific members of the inhibins family) provide the highest sensitivity/specificity characteristics as an ovarian cancer diagnostic test (Mol Cell Endocrinol.
2002 May 31; I 91 ( 1 ) :97-103). Measurement of serum TGF-alpha itself was found to have sensitivity up to 70% and specificity of 89% in early stage disease. The growth factor Mesothelin was also found to have diagnostic value but only for late stage disease.
Immunohistochemistry is frequently used to assess the origin of tumor and staging when a pathological tissue sample is available. A few molecular markers have been shown to have diagnostic value in Immunohistochemistry of ovarian cancer, among them Epidermal Growth Factor, p53 and HER-2. P53 expression is much lower at early stage than late stage disease. P53 high expression is more typical or characteristic of invasive serous tumors than of mucinous I 0 tumors. No benign tumors are stained with P53. HER-2 is found in less than 25% of newly diagnosed ovarian cancers. Ovarian cancer of type granulosa cell tumor has in general better prognosis with late relapse and/or metastasis formation. However, about 50% of patients still die within 20 years of diagnosis. In this specific tumor type, immunohistochemistry staining of estrogen receptor beta (ERb) and proliferating cell nuclear antigen (PCNA) showed that loss of ERb expression and high PCNA expression, characterized a subgroup of granulosa cell tumors with a worse outcome (Histopathology. 2003 Sep;43(3):254-62). Survivin expression was also shown to be correlated to tumor grade, histologic type and mutant p53 but actual correlation to survival is questionable (Mod Pathol. 2004 Feb;17(2):264) Many other markers have been tested over the years for ovarian cancer detection. Some markers have shown only limited value while others are still under investigation. Among them are TPA and TPS, two cytokeratins whose inclusion in a panel with CA-125 resulted in diagnoses with sensitivity up to 93% and specificity up to 98%. LPA -lysophosphatidic acid was a very promising marker with one study demonstrating 98% sensitivity and 90% specificity.
However, this marker is very unstable and requires quick processing and freezing of plasma, and 25. therefore has limited usage.
As previously described, no single marker has been shown to be sufficiently sensitive or specific to contribute to the diagnosis of ovarian cancer. Therefore combinations of markers in panel are being tested. Usually CA-125 is one of the panel members. The best performing panel combinations so far have been CA-125 with CA 15-3 with sensitivity of 93% and specificity of 93%, CA-125 with CEA (which has very little sensitivity by itself with specificity of 93% and specificity of 93%, and CA-125 with TAG-72 and CA 15-3 where specificity becomes 95% but sensitivity is diminished (Dis Markers. 2004;20(2):53-70).
SUMMARY OF THE INVENTION
The background art does not teach or suggest markers for ovarian cancer that are sufficiently sens hive and/or accurate, alone or in combination.
The present invention overcomes these deficiencies of the background art by providing novel markers for ovarian cancer that are both sensitive and accurate. These markers are differentially expressed and preferably overexpressed in ovarian cancer specifically, as opposed to normal ovarian tissue. The measurement of these markers, alone or in combination, in patient (biological) samples provides information that the diagnostician can correlate with a probable diagnosis of ovarian cancer. The markers of the present invention, alone or in combination, show a high degree of differential detection between ovarian cancer and norrcancerous states.
According to preferred embodiments of the present invention, examples of suitable biological samples which may optionally be used with preferred embodiments of the present invention include but are not limited to blood, serum, plasma, blood cells, urine, sputum, saliva, stool, spinal fluid or CSF, lymph fluid, the external secretions of the skin, respiratory, intestinal, and genitourinary tracts, tears, milk, neuronal tissue, ovarian tissue, any human organ or tissue, including any tumor or normal tissue, any sample obtained by lavage (for example of the bronchial system or of the female reproductive system), and also samples of in vivo cell culture constituents. In a preferred embodiment, the biological sample comprises ovarian tissue and/or a serum sample and/or a urine sample and/or secretions or other samples from the female reproductive system and/or any other tissue or liquid sample. The sample can optionally be diluted with a suitable eluant before contacting the sample to an antibody and/or performing any other diagnostic assay.
Information given in the text with regard to cellular localization was determined according to four different software programs: (i) tmhmm (from Center for Biological Sequence Analysis, Technical University of Denmark DTU, http://www.ebs.dtu.dk/services/TMHMM/TMHMM2.Ob.guide.php) or (ii) tmpred (from EMBnet, maintained by the ISREC Bionfonnatics group and the LICK Information Technology Office, Ludwig Institute for Cancer Research, Swiss Institute of Bioinfornatics, http://www.ch.embnet.org/software/TMPRED form.html) for transmembrane region prediction; (iii) signalp hmm or (iv) signalp nn (both from Center for Biological Sequence 5 Analysis, Technical University of Denmark DTU, http://www.cbs.dtu.dk/services/SignalP/background/prediction.php) for signal peptide prediction. The terms "signalp hmm" and "signalp nn" refer to two modes of operation for the program SignalP: hmm refers to Hidden Markov Model, while nn refers to neural networks.
Localization was also determined through manual inspection of known protein localization and/or gene structure, and the use of heuristics by the individual inventor.
In some cases for the manual inspection of cellular localization prediction inventors used the ProLoc computational platform [Einat Hazkani-Covo, Erez Levanon, Galit Rotman, Dan Graur and Amit Novik;
(2004) "Evolution of multicellularity in metazoa: comparative analysis of the subcellular localization of proteins in Saccharomyces, Drosophila and Caenorhabditis."
Cell Biology IS International 2004;28(3):171-8.], which predicts protein localization based on various parameters including, protein domains (e.g., prediction of trans-membranous regions and localization thereof within the protein), pI, protein length, amino acid composition, homology to pre-annotated proteins, recognition of sequence patterns which direct the protein to a certain organelle (such as, nuclear localization signal, NLS, mitochondria localization signal), signal peptide and anchor modeling and using unique domains from Pfam that are specific to a single compartment.
Information is given in the text with regard to SNPs (single nucleotide polymorphisms).
A description of the abbreviations is as follows. "T - > C", for example, means that the SNP
results in a change at the position given in the table from T to C. Similarly, "M - > Q", for example, means that the SNP has caused a change in the corresponding amino acid sequence, from methionine (M) to glutamine (Q). If, in place of a letter at the right hand side for the nucleotide sequence SNP, there is a space, it indicates that a frameshift has occurred. A
frameshift may also be indicated with a hyphen (-). A stop codon is indicated with an asterisk at the right hand side (*). As part of the description of an SNP, a comment may be found in parentheses after the above description of the SNP itself. This comment may include an FTId, which is an identifier to a SwissProt entry that was created with the indicated SNP. An FTId is a unique and stable feature identifier, which allows construction of links directly from position specific annotation in the feature table to specialized protein-related databases. The FTId is always the last component of a feature in the description field, as follows:
FTId=XXX number, in which XXX is the 3-letter code for the specific feature key, separated by an underscore from a Crdigit number. In the table of the amino acid mutations of the wild type proteins of the selected splice variants of the invention, the header of the first column is "SNP positions) on amino acid sequence", representing a position of a known mutation on amino acid sequence.
SNPs may optionally be used as diagnostic markers according to the present invention, alone or in combination with one or more other SNPs and/or any other diagnostic marker.
Preferred embodiments of the present invention comprise such SNPs, including but not limited to novel SNPs on the known (WT or wild type) protein sequences given below, as well as novel nucleic acid andlor amino acid sequences formed through such SNPs, and/or any SNP on a variant amino acid and/or nucleic acid sequence described herein.
Information given in the text with regard to the Homology to the known proteins was determined by SmitIrWaterman version 5.1.2 using special (non default) parameters as follows:
-model=sw.model -GAPEXT=0 -GAPOP=100.0 -MATRIX=blosuml 00 Information is given with regard to overexpression of a cluster in cancer based on ESTs.
A key to the p values with regard to the analysis of such overexpression is as follows:
- librar~based statistics: P-value without including the level of expression in cell-lines (P1) - library based statistics: P-value including the level of expression in cell-lines (P2) - EST clone statistics: P-value without including the level of expression in cell- lines (SP 1 ) - EST clone statistics: predicted overexpression ratio without including the level of expression in cell-lines (R3) - EST clone statistics: P-value including the level of expression in cell-lines (SP2) - EST clone statistics: predicted overexpression ratio including the level of expression in cell-lines (R4) Library based statistics refer to statistics over an entire library, while EST
clone statistics refer to expression only for ESTs from a particular tissue or cancer.
Information is given with regard to overexpression of a cluster in cancer based on microarrays. As a microarray reference, in the specific segment paragraphs, the unabbreviated tissue name was used as the reference to the type of chip for which expression was measured.
There are two types of microarray results: those from microarrays prepared according to a design by the present inventors, for which the microarray fabrication procedure is described in detail in Materials and Experimental Procedures section herein; and those results from microarrays using Affymetrix technology. As a microarray reference, in the specific segment paragraphs, the unabbreviated tissue name was used as the reference to the type of chip for which expression was measured. For microarrays prepared according to a design by the present I S inventors, the probe name begins with the name of the cluster (gene), followed by an identifying number. These probes are listed below with their respective sequences.
>H61775 0 11 0 CCCCAGCTTTTATAGAGCGGCCCAAGGAAGAATATTTCCAAGAAGTAGGG
>HSAPHOL 0 I1 0 GGAACATTCTGGATCTGACCCTCCCAGTCTCATCTCCTGACCCTCCCACT
>HUMGRPSE 0 0 16630 GCTGATATGGAAGTTGGGGAATCTGAATTGCCAGAGAATCTTGGGAAGAG
>HUMGRPSE 0 2 0 TCTCATAGAAGCAAAGGAGAACAGAAACCACCAGCCACCTCAACCCAAGG
>D56406 0 5 0 TCTGACTTTTACGGACTTGGCTTGTTAGAAGGCTGAAAGATGATGGCAGG
>M77904 0 8 0 AGTCTGTGTTTGAGGGTGAAGGCTCAGCAACCCTGATGTCTGCCAACTAC
>Z25299 0 3 0 AACTCTGGCACCTTGGGCTGTGGAAGGCTCTGGAAAGTCCTTCAAAGCTG
>Z44808 0 8 0 AAAAGCATGAGTTTCTGACCAGCGTTCTGGACGCGCTGTCCACGGACATG
>Z44808 0 0 72347 ATGTTCTTAGGAGGCAAGCCAGGAGAAGCCGGGTCTGACTTTTCAGCTCA
>Z44808 0 0 72349 TCCTCCAGACCCAAAGCCACAACCCATCGCAAGTCAAGAACACTTTCCAG
>S67314 0 0 741 CACAGAGCCAGGATGTTCTTCTGACCTCAGTATCTACTCCAGCTCCAGCT
>S67314 0 0 744 TGGCATGCTGGAACATGGACTCTAGCTAGCAAGAAGGGCTCAAGGAGGTG
>Z39337 0 0 66755 GCAGGGGTTAAAAGGACGTTCCAGAAGCATCTGGGGACAGAACCAGCCTC
>Z39337 0 9 0 TAATAAACGCAGCGACGTGAGGGTCCTGATTCTCCCTGGTTTTACCCCAG
>HUMPHOSLIP 0 0 18458 AAGGAAGCAGGACCAGTGGATGTGAGGCGTGGTCGAAGAACAACAGAAAG
>HUMPHOSLIP 0 0 18487 ACAGGGGCCAGATGGTGACCCATGACCCAGCCTAAAAGGCAGCCAGAGGG
>M78530 0 6 0 CTTCCTACACACATCTAGACGTTCAAGTTTGCAAATCAGTTTTTAGCAAG
>HSMUC1A 0 37 0 AAAAGGAGACTTCGGCTACCCAGAGAAGTTCAGTGCCCAGCTCTACTGAG
>HSMUC1A 0 0 11364 AAAGGCTGGCATAGGGGGAGGTTTCCCAGGTAGAAGAAGAAGTGTCAGCA
>HSMUC 1 A 0 0 11365 AATTAACCCTTTGAGAGCTGGCCAGGACTCTGGACTGATTACCCCAGCCT
Oligonucleotide microarray results taken from Affymetrix data were from chips available from Affymetrix Inc, Santa Clara, CA, USA (see for example data regarding the Human Genome U133 (HG-U133) Set at www.affymetrix.com/products/arrays/specific/hgul33.affx; GeneChip Human Genome 2.0 Array at www.affymetrix.com/products/arrays/specific/hgu133av2.affx; and Human Genome U133 Plus 2.0 Array at www.affymetrix.com/products/arrays/specific/hgu I 33plus.affx). The probe names follow the Affymetrix naming convention. The data is available from NCBI Gene Expression Omnibus (see www.ncbi.nlm.nih.gov/projects/geo/ and Edgar et al, Nucleic Acids Research, 2002, Vol.
30, No. 1 207-210). The dataset (including results) is available from www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE 1133 for the Series GSE 1133 database (published on March 2004); a reference to these results is as follows: Su et al (Proc Natl Acad Sci U S A. 2004 Apr 20;1 O 1 ( I 6):6062-7. Epub 2004 Apr 09).
The following list of abbreviations for tissues was used in the TAA
histograms. The term l0 "TAA" stands for "Tumor Associated Antigen", and the TAA histograms, given in the text, represent the cancerous tissue expression pattern as predicted by the biomarkers selection engine, as described in detail in examples 1-5 below (the first word is the abbreviation while the second word is the full name):
("BONE", "bone");
("COL", "colon");
("EPI", "epithelial");
("GEN", "general");
("LIVER", "liver");
(~~L~~~~ ~~lung~~)~
("LYMPH", "lymph nodes");
("MARROW", "bone marrow");
("OVA", "ovary");
("PANCREAS", "pancreas");
("PRO", "prostate");
("STOMACH", "stomach");
("TCELL", "T cells");
("THYROID", "Thyroid");
("MAM", "breast");
(..BRAIN.,, ..brain");
("UTERUS", "uterus");
("SKIN", "skin");
("KIDNEY", "kidney");
("MUSCLE", "muscle");
("ADREN", "adrenal");
("HEAD", "head and neck");
5 ("BLADDER", "bladder");
It should be noted that the terms "segment", "seg" and "node" are used interchangeably in reference to nucleic acid sequences of the present invention; they refer to portions of nucleic 10 acid sequences that were shown to have one or more properties as described below. They are also the building blocks that were used to construct complete nucleic acid sequences as described in greater detail below. Optionally and preferably, they are examples of oligonucleotides which are embodiments of the present invention, for example as amplicons, hybridization units and/or from which primers and/or complementary oligonucleotides may optionally be derived, and/or for any other use.
Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this invention belongs. The following references provide one of skill with a general definition of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed.
1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988);
The Glossary of Genetics, Sth Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). All of these are hereby incorporated by reference as if fully set forth herein. As used herein, the following terms have the meanings ascribed to them unless specified otherwise.
As used herein the phrase "ovarian cancer" refers to cancers of the ovary including but not limited to Ovarian epithelial tumors (serous, mucinous, endometroid, clear cell, and Brenner tumor), ovarian germ-cell tumors, (teratoma, dysgerminoma, endodermal sinus tumor, and embryonal carcinoma) and ovarian stromal tumors (originating from granulosa, theca, Sertoli, Leydig, and collagerrproducing stromal cells).
m The teen "marker" in the context of the present invention refers to a nucleic acid fragment, a peptide, or a polypeptide, which is differentially present in a sample taken from subjects (patients) having ovarian cancer as compared to a comparable sample taken from subjects who do not have ovarian cancer.
The phrase "differentially present" refers to differences in the quantity of a marker present in a sample taken from patients having ovarian cancer as compared to a comparable sample taken from patients who do not have ovarian cancer. For example, a nucleic acid fragment may optionally be differentially present between the two samples if the amount of the nucleic acid fragment in one sample is significantly different from the amount of the nucleic acid fragment in the other sample, for example as measured by hybridization andlor NAT-based assays. A polypeptide is differentially present between the two samples if the amount of the polypeptide in one sample is significantly different from the amount of the polypeptide in the other sample. It should be noted that if the marker is detectable in one sample and not detectable in the other, then such a marker can be considered to be differentially present.
As used herein the phrase "diagnostic" means identifying the presence or nature of a pathologic condition. Diagnostic methods differ in their sensitivity and specificity. The "sensitivity" of a diagnostic assay is the percentage of diseased individuals who test positive (percent of "true positives"). Diseased individuals not detected by the assay are "false negatives." Subjects who are not diseased and who test negative in the assay are termed "true negatives." The "specificity" of a diagnostic assay is I minus the false positive rate, where the "false positive" rate is defined as the proportion of those without the disease who test positive.
While a particular diagnostic method may not provide a definitive diagnosis of a condition, it suffices if the method provides a positive indication that aids in diagnosis.
As used herein the phrase "diagnosing" refers to classifying a disease or a symptom, determining a severity of the disease, monitoxing disease progression, forecasting an outcome of a disease and/or prospects of recovery. The term "detecting" may also optionally encompass any of the above.
Diagnosis of a disease according to the present invention can be effected by determining a level of a polynucleotide or a polypeptide of the present invention in a biological sample obtained from the subject, wherein the level determined can be correlated with predisposition to, or presence or absence of the disease. It should be noted that a "biological sample obtained from the subject" may also optionally comprise a sample that has not been physically removed from the subject, as described in greater detail below.
As used herein, the term "level" refers to expression levels of RNA and/or protein or to DNA copy number of a marker of the present invention.
Typically the level of the marker in a biological sample obtained from the subject is different (i.e., increased or decreased) from the level of the same variant in a similar sample obtained from a healthy individual (examples of biological samples are described herein).
Numerous well known tissue or fluid collection methods can be utilized to collect the biological sample from the subject in order to determine the level of DNA, RNA
and/or polypeptide of the variant of interest in the subject.
Examples include, but are not limited to; fine needle biopsy, needle biopsy, core needle biopsy and surgical biopsy (e.g., brain biopsy), and lavage. Regardless of the procedure employed, once a biopsy/sample is obtained the level of the variant can be determined and a diagnosis can thus be made.
Determining the level of the same variant in normal tissues of the same origin is preferably effected along-side to detect an elevated expression and/or amplification and/or a decreased expression, of the variant as opposed to the normal tissues.
A "test amount" of a marker refers to an amount of a marker in a subject's sample that is consistent with a diagnosis of ovarian cancer. A test amount can be either in absolute amount (e.g., microgram/ml) or a relative amount (e.g., relative intensity of signals).
A "control amount" of a marker can be any amount or a range of amounts to be compared against a test amount of a marker. For example, a control amount of a marker can be the amount of a marker in a patient with ovarian cancer or a person without ovarian cancer. A
control amount can be either in absolute amount (e.g., microgram/ml) or a relative amount (e.g., relative intensity of signals).
"Detect" refers to identifying the presence, absence or amount of the object to be detected.
A "label" includes any moiety or item detectable by spectroscopic, photo chemical, biochemical, immunochemical, or chemical means. For example, useful labels include 32p, 3sS, fluorescent dyes, electrorrdense reagents, enzymes (e.g., as commonly used in an ELISA), biotin-streptavadin, dioxigenin, haptens and proteins for which antisera or monoclonal antibodies are available, or nucleic acid molecules with a sequence complementary to a target.
The label often generates a measurable signal, such as a radioactive, chromogenic, or fluorescent signal, that can be used to quantify the amount of bound label in a sample. The label can be incorporated in or attached to a primer or probe either covalently, or through ionic, van der Waals or hydrogen bonds, e.g., incorporation of radioactive nucleotides, or biotinylated nucleotides that are recognized by streptavadin. The label may be directly or indirectly detectable. Indirect detection can involve the binding of a second label to the first label, directly or indirectly. For example, the label can be the ligand of a binding partner, such as biotin, which is a binding partner for streptavadin, or a nucleotide sequence, which is the binding partner for a complementary sequence, to which it can specifically hybridize. The binding partner may itself be directly detectable, for example, an antibody may be itself labeled with a fluorescent molecule. The binding partner also may be indirectly detectable, for example, a nucleic acid having a complementary nucleotide sequence can be a part of a branched DNA
molecule that is in turn detectable through hybridization with other labeled nucleic acid molecules (see, e.g., P.
D. Fahrlander and A. Klausner, Bio/Technology 6:1165 (1988)). Quantitation of the signal is achieved by, e.g., scintillation counting, densitometry, or flow cytometry.
Exemplary detectable labels, optionally and preferably for use with immunoassays, include but are not limited to magnetic beads, fluorescent dyes, radiolabels, enzymes (e.g., horse radish peroxide, alkaline phosphatase and others commonly used in an ELISA), and calorimetric labels such as colloidal gold or colored glass or plastic beads.
Alternatively, the marker in the sample can be detected using an indirect assay, wherein, for example, a second, labeled antibody is used to detect bound marker-specific antibody, and/or in a competition or inhibition assay wherein, for example, a monoclonal antibody which binds to a distinct epitope of the marker are incubated simultaneously with the mixture.
"Immunoassay" is an assay that uses an antibody to specifically bind an antigen. The immunoassay is characterized by the use of specific binding properties of a particular antibody to isolate, target, andlor quantify the antigen.
The phrase "specifically (or selectively) binds" to an antibody or "specifically (or selectively) immunoreactive with," when referring to a protein or peptide (or other epitope), refers to a binding reaction that is determinative of the presence of the protein in a heterogeneous population of proteins and other biologics. Thus, under designated immunoassay conditions, the specified antibodies bind to a particular protein at least two times greater than the background (non-specific signal) and do not substantially bind in a significant amount to other proteins present in the sample. Specific binding to an antibody under such conditions may require an antibody that is selected for its specificity for a particular protein. For example, polyclonal antibodies raised to seminal basic protein from specific species such as rat, mouse, or human can be selected to obtain only those polyclonal antibodies that are specifically immunoreactive with seminal basic protein and not with other proteins, except for polymorphic variants and alleles of seminal basic protein. This selection may be achieved by subtracting out antibodies that cross-react with seminal basic protein molecules from other species. A variety of immunoassay formats may be used to select antibodies specifically immunoreactive with a particular protein. For example, solid-phase ELISA immunoassays are routinely used to select antibodies specifically immunoreactive with a protein (see, e.g., Harlow &
Lane, Antibodies, A
Laboratory Manual (1988), for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity). Typically a specific or selective reaction will be at least twice background signal or noise and more typically more than 10 to 100 times background.
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~,...
~T'ranscript Name ;u~ a.. ,. ..
z H571775 T21 .
a nucleic acid sequence comprising a sequence in the table below:
'ur .~ ~ ' a,.
>,'~Segme°n~C Name -~ ~ , ø._._. x H617?5 node 2 H61775 node 4 H61775 node 6 H61775 node 8 H61775 node 0 H61775 node 5 According to preferred embodiments of the present invent ion, there is provided an isolated polypeptide comprising an amino acid sequence in the table below amino acid sequence comprising a sequence in the table below:
Prot~ln~Naane ~:'~ ~~'~
- ~ -~
~ : ~~ v ~. ..~, ~z ..s,::%~ :m."~..,.~-.'-~;
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
Transcript~ame - ~' ~
~
~~, ~ ~~- ~_.
L ~~.~
10 a nucleic acid sequence comprising a sequence in the table below:
Se ent Name ~
~$
~' ''~~s ~~~
HUMCEA PEA 1 node 0 HUMCEA PEA 1 node 2 HUMCEA PEA 1 node 11 HUMCEA PEA 1 node 12 HUMCEA PEA 1 node 31 HUMCEA PEA I
node node node node node node node node node HUMCEA PEA node9 I
HUMCEA PEA node10 HUMCEA PEA node15 HUMCEA PEA node16 HUMCEA PEA node17 I
HUMCEA PEA node18 HUMCEA PEA node19 HUMCEA PEA node20 HUMCEA PEA node21 I
HUMCEA PEA node22 HUMCEA PEA node23 HUMCEA PEA node24 HUMCEA PEA node27 HUMCEA PEA node29 HUMCEA PEA node30 HUMCEA PEA node33 HUMCEA PEA node34 HUMCEA PEA node35 HUMCEA PEA node HUMCEA PEA node50 HUMCEA PEA node51 HUMCEA PEA node 56 HUMCEA PEA node 57 I
HUMCEA PEA node 58 HUMCEA PEA node 60 HUMCEA PEA node 61 HUMCEA PEA node 62 HUMCEA PEA-1node 64 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
w,-.~ . ~,.
~.>.. ,.~ w.
~ . ~.. ,Y,.:.~ . ~~ -.~., (~'~~ x ~-..v~,.. ~~orres ondm . Traris~n ~ s s, :
.- .
Protem~Name~ ~x , _, .., ~ ~- ..~
~_~ C~' .:- 9 ' , v. ~
s;~~~ , ' ~
'' .a3:..,.,<. ~
-..a , "
-~,~ , v ~ ..," ~
y~ W"-v~ ,.
~" a:''.~"a=;: ....
: ....
~ sY,,.-x~-"
x,;.
t ~r~ a.~ ": .,~...,~'~:.;-_:~
>. rc.
~
~~
- > ... ':. ~ .,~..~:~".:.,.",.._ ~ ~...~,<> x. _,>a"5-~,. ~.:~-~"~;;;
M
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~'zan~crt'~1~1~~'rne~~~~~~~.
s _ ~
~'~
~
~
'~ ..:..
, .
:
~
a nucleic acid sequence comprising a sequence in the table below:
~Segnient~Narine' '' "'_'~b ,. ~"-HUMEDF PEA 2node6 1-IUMEDF 2node11 PEA
HUMEDF PEA 2nodet 8 H UMEDF 2node19 PEA
HUMEDF PEA 2node22 I-IUMEDF 2node2 PEA
HUMEDF PEA 2node8 I-IUMEDF 2node20 PEA
According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
.,.;e;?';',.~~~' a; ,.:;... ",~'~~.a<-..~.. . , . ~" w~~
..~r,w ...,., a.. .~3""..- .~.~.;....~~'-, ,~a7~ '.." s. .. a. "~ '"~ ,.,~.r~
~L'rotena Name~~~~= -.:~ ~..- , ." -: ~-..r:..f.:.
~.-3 ~ CoiTes on_d~n - ~z~nscr~ t a ~.~v ~;..,: ,;~ .- ":.... :,>. .... ~ ~ o.~ ~..:.,"".
: t ~: ~R
~ a, ~- ~
~? ~
, MY a h 4 ' :~
~W
~
-.
Y
i .."",...; ,..
" ~.
,.. ,. ,~.C
~~m.,.. < ",.w, W~a.
, : s, ..., ~
~,. ~....,.
.,. .., .
,.,.. 7 ..,..
.,.
.
, , ..
.
.
r ~
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below andlor:
s~~': ~ ~. ~r ~~ ~'~~Cl 1~~a?T~~ a~ ~
~.- :,- ~- ~~ , a~
HSAPHOL T10 ~~
HSAPHOL TS
a nucleic acid sequence comprising a sequence in the table below:
~ ~-s ~a~--,_ ~ ~s:.:.' .n.-...-,~. x~ .s m -;-:
Segment Name.
-''~
HSAPHOL node 1 l Y
I-ISAPHOL node 13 HSAPHOL node 15 HSAPHOL node 19 HSAPHOL node 2 HSAPI-IOL node 21 HSAPHOL node 23 HSAPHOL node 26 HSAPHOL node 28 HSAPHOL node 38 HSAPHOL node 40 HSAPHOL node 42 HSAPHOL node 16 HSAPHOL node 25 HSAPHOL node 34 HSAPHOL node 35 HSAPHOL node 36 HSAPHOL node 41 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
r ~1 a~e,w 3 ~~
2o HSAPHOL PS
~HSAPHOL P8 According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
,.m ~ ~ ~ ~--_ °~ f Traris rlpt Name ~ .
y, z ~ ~ w~ . a ;' .'~:._.. r . ~ 2.~.,..?. ~ :es a nucleic acid sequence comprising a sequence in the table below:
Sgt~~en~lame m b ~ y T1088 I ll PEA node node node node node node node node-According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
a ~aa a '~,~ t ~.Y~' ':~ ~t s,~' ~ ~ ~s "' ~P~rotem Name~~~~ ~ F~~ ~~~ ~~Y, , ~.
~. x,7 - X-~..,v ..,~-..»~ .~°.~,.,r... <s~'.e. '~.- , . ,a ~
.~ :_ss T10888 PEA_1 P6 According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~~~ ~r .~
~ ~ ~r scr~pt ~ .~ ~w Nam e ~
~ ~ ~
_ .".R~. ~..a ~,.~.-,~
~ ~ _z ~,. ,.a '.~ . ",,m",~a.
~ m~ , .~,s l a nucleic acid sequence comprising a sequence in the table below:
~~S~e ~i~~it am ~ ~~~~ -'_' ~ ~ ~ ~ x ~,.t~a ~3',.X~a g a F, ~" i~~ na,c ~. A7,'~.
~ z~, . . ? " ~4.. , F~,r,.,B~'I~.lhf,r HSECADH node 0 I-ISECADH node 14 HSECADH node 15 HSECADH node 21 HSECADH node 22 HSECADH node 25 HSECADH node 26 HSECADH node 48 HSECADH node 52 HSECADH node 53 HSECADH node 54 HSECADH node 57 HSECADH node 60 HSECADH node 62 HSECADH node 63 HSECADH node 7 HSECADH node 1 HSECADH node 11 HSECADH node 12 HSECADH node 17 HSECADH node 18 HSECADH node 19 HSECADH node 3 HSECADH node HSECADH node HSECADH node HSECADH node 55 HSECADH node 56 HSECADH node HSECADH node 59 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~~ iltein~Nam~
., a -_s ~ a": a'~.R :. . zoo .
HSECADH P l 5 According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~" ~
Transcript Name- ~--~-~' ~-~ ~ ~-y HUMGRPSE TS
a nucleic acid sequence comprising a sequence in the table below:
~~ Se~gm~e~'nt~~ame a ~~
W
E
rv HUMGRPSE node 0 HUMGRPSE node .
HUMGRPSE node 8 HUMGRPSE node 3 HUMGRPSE node 7 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
Protern~Name~ _ ~ ~ -~ ,~~ ~~
HUMGRPSE PS
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
y Transcri t Na ne p l ~.. ~ _.
R 11723 I T l 7 PEA
PEA
a nucleic acid sequence comprising a sequence in the table below:
~'~.Segrnent~I~am~~~
~~_ p. ~
~_ ~:. .
..~:~, ~F~o._ T: .-.~
-. ., _ W~
811723 PEA l 13 node node node node node node node node node node node node node node node node node 811723 PEA_1 node 811723 PEA node29 R 1 1723PEA node3 R 1 1723PEA node30 R 11723 PEA node4 811723 PEA node5 811723 PEA node6 811723 PEA node7 I
811723 PEA node8 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~,Protem~N~xn~ '~'~ ~~ ~~"xr ~ R
~~:: ~:~,a ,,~_..,._.....:~»..
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~ ~ ~ ~~ . x~, ~~anscri t~~Tama ~~ ~ri ~''~" ~w~.~=g ~ ~ ~,~~ .~
_ ~.~ _~
D56406 PEA 1 T3 ~
a nucleic acid sequence comprising a sequence in the table below:
a, .. . - ...,,.
, a Segment Nalne ,:
a;
~-.
v '..~
a"~~
D56406 PEA node0 I
D56406 PEA node13 D56406 PEA node11 D56406 PEA node2 D56406 PEA node3 D56406 PEA node5 D56406 PEA node6 I
D56406 PEA node7 DS6406 PEA node8 I
D56406_PEA_1 node9 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
rCf>rf31n1'~t~llle~ '~-~~ _ :~, a .,~ r<
According to preferred embodiments ofthe present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
T ~scz~p Name ~ ~ ~ , ~, _, H53393 PEA~1 T9 a nucleic acid sequence comprising a sequence in the table below:
,_ ~ .Segment Name ~ ~
s ~...m~
. E.~
axe~~.>.~
~ ~
x -'.
v ~e H53393 PEA node 0 I
H53393 PEA node 10 H53393 PEA node 12 H53393 PEA node 13 I
H53393 PEA node 1 S
I
H53393 PEA node 17 H53393 PEA node 19 I
H53393 PEA node 23 I
H53393 PEA node 24 H53393 PEA node 25 H53393 PEA node 29 I
H53393 PEA node 4 H53393 PEA node 6 H53393 PEA node 8 I
H53393 PEA node 21 H53393 PEA node 22 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
" .. .., ; -:~ w~~;'m'.o.,. ~1' . .. _~~~
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below andlor:
~~TraiascriptNat~e~' '~ ~ ~ v~~~~.
;~ ~ . ~- ; ~ _ -~-a a _ ~. _ .. , -~ ~f _ ,~-.~ ~.
a nucleic acid sequence comprising a sequence in the table below:
~~Segment~Name~-~
~ ..~~~~
-~~ '--.~' ,.
~
HSU40434 PEA Inode l HSU40434 PEA 1node 16 HSU40434 PEA Inode 30 HSU40434 PEA Inode 32 HSU40434 PEA Inode 57 HSU40434 PEA Inode 0 HSU40434 PEA 1node 10 HSU40434 PEA 1node 13 HSU40434 PEA 1node 18 HSU40434 PEA 1node 2 HSU40434 PEA 1node 20 HSU40434 PEA 1node 21 HSU40434 PEA 1node 23 HSU40434 PEA 1node 24 HSU40434 PEA 1node 26 HSU40434 PEA 1node 28 HSU40434 PEA 1node 3 HSU40434 PEA 1node 35 HSU40434 PEA 1node 36 HSU40434 PEA 1node 37 HSU40434 PEA 1node 38 I-ISU40434 PEA node 39 I
HSU40434 PEA node 40 HSU40434 PEA node 41 HSU40434 PEA node 42 HSU40434 PEA node 43 I
HSU40434 PEA node 44 HSU40434 PEA node 47 I
HSU40434 PEA node 48 HSU40434 PEA node 51 HSU40434 PEA node 52 HSU40434 PEA node 53 HSU40434 PEA node 54 I
HSU40434 PEA node 56 HSU40434 PEA node 7 HSU40434 PEA-1 node-8 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
PrOt~81r1~ ~ ~ '~~~;~~~ ~- " S
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~' 'I~rans ~t TT~~ ne ~z x, .~~,~ ~ _:
a nucleic acid sequence comprising a sequence in the table below:
~_ ~~ ~ gment~larne~ ~ ~ ~ ~~ ~ ~ ~ ~=, ~, ~ø_ -~ w....~..__.,.
M77904 node 0 M77904 node 11 M77904 node 12 M77904 node 14 M77904 node 15 M77904 node 17 M77904 node 2 M77904 node 21 M77904 node 23 M77904 node 24 M77904 node 27 M77904 node 28 M77904 node 4 M77904 node 6 M77904 node 7 M77904 node 8 M77904 node 9 M77904 node 19 M77904 node 22 M77904 node 25 M77904 node 26 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
Protein e ~~ ' ~ : ~ ~ '~m~.
Nam ~~ ~~
s ~ a ..~
P , -; "~: _ -~i=Tv'- ;~--.
'',n_aa;,. , '," e' -...
PS
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~, ~
'Txanscnpt Name ~_ ~ ~ >~~ ~ ~ ri~
_,.,. ~, ~ "~ r,p.
s' .. ;':~ ~":, .«.., Wit' 's '~" , ~ ''~;_,' .~,a g a nucleic acid sequence comprising a sequence in the table below:
~egment~
am ~ x T
~~ ~ ~ "
~m.
w _..~ r w_~,.;~
'~z :'8~
:~ ~ 'a~s.~:y'~i~~
~,'~ . ,.
~ ~ .z:"
225299 PEA 2 node 20 225299 PEA 2 node 21 225299 PEA 2 node 23 225299 PEA 2 node 24 225299 PEA 2 node 8 225299 PEA 2 node 12 2 node 13 225299 PEA 2 node 14 2 node 17 225299 PEA 2 node 18 225299 PEA 2 node 19 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
Pr_oteinNa~ie ~~ ~~.~~~~ ~~_~r ~, ' g~ ~, ='~
Z25299~ PEA 2 P2 According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
TIaIi~Cl'l~t NalTte~° '~x~' ~ a, T39971 T1"0 a nucleic acid sequence comprising a sequence in the table below:
a en ame ~~
~'~..~' ~
' ~
,~
.;..
,:
T39971node0 T39971node18 T39971node21 T39971node22 T39971node23 T39971node31 T39971node33 9971 node7 T39971node1 T39971node10 T39971node11 T39971node12 T39971node15 T39971node16 T39971node17 T39971node26 T39971node27 T39971node28 T39971node29 T39971node3 T39971node30 T39971node34 T39971node35 T39971node36 T39971node4 T39971node5 T39971node8 T39971node9 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
Protein Name ~ ~~~i ~'~~ °" °~ ~ ~ ~:
f A~~~~~.,n.~ o According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~ a s...
Transcript Name~~~- ~ ~~m ~~ ~.
244808 .PEA 1 T5 a nucleic acid sequence comprising a sequence in the table below:
~Se~grnent~Name 244808PEAI node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808 1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node II
244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA1 node 244808PEA 1 node 35 244808PEA 1 node 39 244808PEA node 4 I
244808PEA node 6 44808PEA-1node 8 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
Paz ~P~,rote~n Nar~n'e ~,~
_s ~
~k,..;;...~~,~ . r,.~.''.~_ . ~....,~... .~" ~ ~y,,r '~:, According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~TrdIISCrI~t ~8 "' "s~~"w a nucleic acid sequence comprising a sequence in the table below:
Segra~ent Name~~
' ~
-P' T
g -:
.~k~
.
bh~.,...<.~"K9 ~,.'~.~,'.~5r ' S67314 node 0 PEA
567314PEA node 11 I
567314PEA node 13 567314PEA node 15 l 567314PEA 1 node 17 567314PEA node 4 S67314PEA node 10 567314PEA node 3 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
otem Name ~ ~
~ ~' ~~ ~ z ~:
' ' ~
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~'P a scnpt~ a"nie~°~ '~~"~~c ~x~ ~~ ~:u ~3 ~~
.~ _~~.-.~ .
239337 PEA t2 PEA 1 T3 239337 PEA 2 PEA_1 T12 a nucleic acid sequence comprising a sequence in the table below:
~~t N~_ tee . ;
~ ~ y ~ a 2 .. ,~T.,'~
_~..~'' ' ~,.i., 239337 PEA PEA node 2 239337 PEA PEA_1node-15 239337 PEA PEA_1node_16 239337 PEA PEA node 18 239337 PEA PEA node 21 239337 PEA PEA-1node 22 239337PEA 2 PEA node 3 239337PEA 2 PEA node 5 239337PEA 2 PEA node 6 239337PEA 2 PEA node 10 239337PEA 2 PEA node 11 239337 2 PEA node 14 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
z,:.xa. .~ ,:; .x ' ~.~, ~ 4'~~:...
~ .~,;;-:3~.~ s .~....'~~ ,x. -:-~ ., sa .
vs , rs ,.::::.-~',";cx;r ,<;,.,a" n ra , ,~;~.,:~t,,,."..w'~
s ,.3",??
r ."'..s:' ., .,?.':'. "~ .l:~c: ~ tea- ~ a ae. ' ,., a ;~ : .,1 Prot~mName~~~.~ ~~. _ _~w_ w~ ~ r ~~ ~~ ,~ ~_~~~.-~ ~' . ~, ~
a ~~
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
Txanscnp aye ..,..~.':.~-o.~.
,~~'f~' s~
~3 ' M Y~
~ E~ i~ ~Hf ~~
PEA
PEA
PEA
PEA
PEA
a nucleic acid sequence comprising a sequence in the table below:
~5entN~'~aine ~~, ~- ~~ ~ ~ ,-u-~:
~w ~..:. ~. _ ~~.,~ .; a HUMPHOSLIP PEA 2 node 0 HUMPHOSLIP 2 node19 PEA
HUMPHOSLIP 2 node34 PEA
HUMPHOSLIP 2 node68 PEA
HUMPHOSLIP 2 node70 PEA
HUMPHOSL1P 2 node75 PEA
HUMPHOSLIP 2 node2 PEA
HUMPHOSLIP 2 node3 PEA
HUMPHOSLIP 2 node HUMPHOSLIP 2 node6 PEA
HUMPHOSLIP 2 node7 PEA
HUMPHOSLIP 2 node8 PEA
HUMPHOSLIP 2 node9 PEA
HUMPHOSLIP 2 nodeI4 PEA
HUMPHOSLIP 2 node15 PEA
HUMPHOSLIP 2 node16 PEA
HUMPHOSLIP 2 node17 PEA
HUMPHOSLIP node23 HUMPHOSLIP node24 HUMPHOSLIP 2 node HUMPHOSLIP 2 node26 PEA
HUMPHOSLIP 2 node PEA node HUMPHOSLIP node33 HUMPHOSLIP node36 HUMPHOSLIP node37 HUMPHOSLIP node39 HUMPHOSLIP node HUMPHOSLIP node HUMPHOSLIP node HUMPHOSLIP 2 node HUMPI-IOSLIP node 45 HUMPHOSLIP PEA node 47 HUMPHOSLIP PEA node 51 HUMPHOSLIP PEA node 52 HUMPHOSLIP PEA node 53 HUMPHOSLIP PEA node 54 HUMPHOSLIP PEA node 55 HUMPHOSLIP PEA node 58 HUMPHOSLIP PEA node 59 HUMPHOSLIP PEA node 60 HUMPHOSLIP PEA node 61 HUMPHOSLIP PEA node 62 HUMPHOSLIP PEA node 63 HUMPHOSLIP PEA node 64 HUMPHOSLIP PEA node 65 HUMPHOSLIP PEA node 66 HUMPHOSLIP PEA node 67 HUMPHOSLIP PEA node 69 HUMPHOSLIP PEA node 71 HUMPHOSLIP PEA node 72 .2 HUMPHOSLIP PEA node 73 HUMPHOSLIP PEA node 74 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
PEA
PEA
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~Tr nsc pt Named ~
a nucleic acid sequence comprising a sequence in the table below:
~Se ent l~Iactae T59832 node 1 T59832 node 7 T59832 node 29 T59832 node 39 T59832 node 2 T59832 node 3 T59832 node 4 T59832 node 5 T59832 node 6 T59832 node 8 T59832 node 9 T59832 node 10 T59832node11 T59832node12 T59832node14 T59832node16 T59832node19 T59832node20 T59832node25 T59832node26 T59832node27 T59832node28 T59832node30 T59832node31 T59832node32 T59832node34 T59832node35 T59832node36 T59832node37 59832 node38 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~ ~~M =
tem N ~~ .~
iie~
o ai s r~' ~ ~
~ ~. w ~~ ~ .~
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~TranSCript Name -.
~3.r ~TA.
Yk ~.
~"
PEA
a nucleic acid sequence comprising a sequence in the table below:
"ms's ~x~~~"''~' a'""~"F
' ~x Segment~~~lame ~~-~ _ ~ ~",.
M
z"
i"' , ~"yet, .F
HSCP2 PEA node 0 HSCP2 PEA node 3 HSCP2 PEA node 6 HSCP2 PEA node 8 HSCP2 PEA node 10 HSCP2 PEA node 14 HSCP2 PEA node 23 HSCP2 PEA node 26 HSCP2 PEA node 29 HSCP2 PEA node 31 HSCP2 PEA node 32 I
HSCP2_PEA node 34 HSCP2 I node 52 ' PEA
HSCP2 1 node 58 PEA
HSCP2 1 node 72 PEA
HSCP2 1 node 73 PEA
HSCP2 1 node 74 PEA
HSCP2 1 node 76 PEA
HSCP2 1 node 78 PEA
HSCP2 1 node 80 PEA
HSCP2 1 node 84 PEA
HSCP2 1 node 4 PEA
HSCP2 1 node 7 PEA
HSCP2 1 node 13 PEA
HSCP2 1 node I S
PEA
HSCP2 1 node 16 PEA
HSCP2 1 node 18 PEA
HSCP2 1 node 20 PEA
HSCP2 1 node 21 PEA
HSCP2 1 node 37 PEA
HSCP2 1 node 38 PEA
HSCP2 1 node 39 PEA
HSCP2 1 node 41 PEA
HSCP2 1 node 42 PEA
HSCP2 1 node 46 PEA
HSCP2 1 node 47 PEA
HSCP2 1 node SO
PEA
HSCP2 1 node 51 PEA
HSCP2 1 node 55 PEA
HSCP2 1 node 56 PEA
HSCP2 1 node 60 PEA
HSCP2 1 node 61 PEA
HSCP2 PEA node67 HSCP2 PEA node68 HSCP2 PEA node69 HSCP2 PEA node70 I-ISCP2 node75 HSCP2 PEA node77 I
HSCP2 PEA node79 HSCP2 PEA node82 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
"'~"'.r'4.'~";~i3 ~ ..a~~
~,!'.4 ~ '~ &
, S ~.
.~
_,~ k~l I k ,. S. , ~r 'i;.",&
-~' ,.~,~;.
t~. ., ~,:,'~~
~. 3i' ;,~ ..,,y>'a:"'e Prorexn Name _ ~ , ~ ~
_ ~ ~~;, ~ .,~ :v~
~ .,; .
"..,...g~
w'~~~.
" x E:
fi~ s ..~-y~.x'~
~'e~ fli, '.W>p _ h~S_ ~' "G ;i~5 A~ ~ ,s , ,t'...
v s ~ ~T..
-' e. ' A....i"~
~ P'~..
.
.~2s~~~
~:,...~:aa ...._:.-zur:
'a-f .,~',.e"a'-' ': ;
' y ~ ~
- . _ k~.
., ~ -n~.
..a -- ~,..ma....e.
~~ ~ ,.,~
r.....,'?~~
.~Tlz _ .
I P4 s I
I
I
I
I
I
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
E ~~ ~ ~~ ~H
Traos tlpt Nam e~
t r~
HUMTEN~~PEA I T4 ( I
I
PEA
PEA
PEA
PEA
a nucleic acid sequence comprising a sequence in the table below:
~~ m~~~
~' ~~ ,~~
~a~ ~ ~,;
S~ e~t~Narne ~: _ T
HUMTEN PEA 1 node 0 HUMTEN PEA 1 node 2 HUMTEN PEA 1 node 5 HUMTEN PEA I node 6 HUMTEN PEA 1 node 11 HUMTEN PEA 1 node 12 HUMTEN PEA 1 node 16 HUMTEN PEA 1 node t9 HUMTEN PEA 1 node 23 HUMTEN PEA 1 node 27 I-IUMTEN 1 node 28 PEA
HUMTEN PEA 1 node 30 HUMTEN PEA 1 node 32 HUMTEN PEA I node 33 I-IUMTEN 1 node 35 PEA
1-IUMTEN l node 38 PEA
HUMTEN PEA 1 node 40 HUMTEN PEA 1 node 42 HUMTEN PEA 1 node 43 HUMTEN PEA 1 node 44 HUMTEN PEA 1 node 45 HUMTEN .PEA1 node 46 HUMTEN PEA 1 node 47 HUMTEN PEA 1 node 49 HUMTEN PEA 1 node 51 HUMTEN PEA 1 node 56 HUMTEN PEA 1 node 65 HUMTEN PEA 1 node 71 HUMTEN PEA 1 node 73 HUMTEN PEA 1 node 76 HUMTEN PEA 1 node 79 HUMTEN PEA 1 node 83 HUMTEN PEA 1 node 89 HUMTEN PEA 1 node 7 HUMTEN PEA 1 node 8 HUMTEN PEA 1 node 9 HUMTEN PEA_1 node_14 HUMTEN PEA 1 node 17 HUMTEN PEA 1 node 21 HUMTEN PEA 1 node 22 HUMTEN PEA 1 node 25 HUMTEN PEA 1 node 36 HUMTEN PEA 1 node 53 HUMTEN PEA 1 node 54 HUMTEN PEA 1 node 57 HUMTEN PEA 1 node 61 HUMTEN PEA 1 node 62 HUMTEN PEA 1 node 67 HUMTEN PEA 1 node 68 HUMTEN PEA 1 node 69 HUMTEN PEA 1 node 70 HUMTEN PEA 1 node 72 HUMTEN PEA 1 node 84 HUMTEN PEA 1 node 85 HUMTEN PEA 1 node 86 HUMTEN PEA 1 node 87 HUMTEN PEA 1 node 88 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
'~"~ a ~~
aid'. ~r ~','t' '~' .'~ ~:-r ~. "
otean ame .. ",: , ~~.w,.~,sr~x"~.4~~--.';~','~;~~2.x:..:~~.._s~-,r~'-~:.,.~;_a":~,~'~'~z..~~b4;,i .5":,h;w:~._.:
HUMTEN PEA PS
I
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~r~'Y"~' ~
;~ .,~ z 2.~Y. .,.
,~~~,,, .
r r~ s ~ . 3s ~T~an~~'rlpt ~'G~~
.' a , . .~
r ~
..
~~
PEA
PEA
a nucleic acid sequence comprising a sequence in the table below:
a, egme~t ~,arn~ ~ ,~ 'r 'c-x ~~c s~
HUMOSTRO PEA 1~ PEA 1 node 0 HUMOSTRO PEA 1 PEA 1 node 10 HUMOSTRO PEA 1 PEA 1 node 16 HUMOSTRO PEA 1 PEA 1 node 23 1-IUMOSTRO 1 PEA node31 I-1UMOSTRO 1 PEA node HUMOSTRO PEA 1 PEA node3 HUMOSTRO PEA 1 PEA node5 HUMOSTRO PEA I PEA node7 HUMOSTRO PEA I PEA node8 HUMOSTRO PEA 1 PEA nodeI S
HUMOSTRO PEA 1 PEA node17 HUMOSTRO PEA I PEA node20 HUMOSTRO PEA 1 PEA node21 HUMOSTRO PEA 1 PEA node22 HUMOSTRO PEA 1 PEA node24 I
HUMOSTRO PEA 1 PEA node26 HUMOSTRO PEA 1 PEA node27 HUMOSTRO PEA I PEA node HUMOSTRO PEA I PEA node HUMOSTRO PEA 1 PEA node30 I
HUMOSTRO PEA I PEA node32 I
HUMOSTRO PEA I PEA node34 HUMOSTRO PEA 1 PEA node36 HUMOSTRO PEA 1 PEA node37 HUMOSTRO PEA I PEA node38 I
HUMOSTRO PEA 1 PEA node39 HUMOSTRO PEA 1 PEA node HUMOSTRO PEA I PEA node HUMOSTRO PEA I PEA node According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~_. ,~
-~:~....> .
':~~ ~ ~, ;.-y -: i ;r .
P otyn Name ,.~.a ~ ;. ~-~ ~. ~,...
.~ ~~ , d ~ . ~
3 .. ~ >!.
~' ,. ~
z '.
~
'.' ~
~...~
.. , a dt :
..,:--8,1 .. d._ y. .~.,0.~"-w '..:~'F.- .: ~~~... ~
,. ;r...kw ~~, .x,a4~& ,.,:~ , ~s,_..
t ~
~
~
HUMOSTRO -, PEA 1 .
PEA
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~Transcnpt Nam '~ ~ ~
~ ~~~':ar"..
f.~ 'k:.~
.. r.~sW
.e~ .CEO
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA_1 a nucleic acid sequence comprising a sequence in the table below:
~~SegrnentName a~l~ ,r'' ~ "~ ~r .'..
_..,:
T46984 PEA node 2 I
T46984 PEA node 4 T46984 PEA node 6 I
T46984 PEA node 12 T46984 PEA node 14 T46984 PEA node 25 T46984 PEA node 29 I
T46984 PEA node 34 I
T46984 PEA node 46 T46984 PEA node 47 I
T46984 PEA node 52 T46984 PEA node 65 I
T46984 PEA node 69 T46984 PEA node 75 T46984 PEA node 86 I
T46984 PEA node 9 T46984 PEA node 13 T46984 PEA node 19 T46984 PEA node 21 T46984 PEA node 22 T46984 PEA node 26 T46984 PEA node 28 node node node node node node node node node node node node node node node node node node node node node node node node node node ~T46984PEA-1 85 node According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
_. ~,. ~
s~. 't y ~ - Y.~
~~ ~_, ~~ =
rote~n Name y~ w. ' ~ , T~.~
~ ..~.~
A~. w ~
.'~f. y'~a.
..~ y~
~ .~~..wT
.~, i,, , R " ~i.yofi~...
'.:
"i- ~ ~
' ''F.'~
'T'L.-Y~'m . ~. x.~.w~'~i' (. ~y '-:
'z'~f Y:
..., _4~.
~,.ra.~
,.,:.e:
A., t.'3dd~-'s-..";".'ki ~ r= ."
~ m '~'~'.~..'~,~,~
2., .t .' I;~."
x' 3..
~ - ~a5 '~,;'v s.~;,.~~-s~aC
, ,.:..A;',#Y;R
I
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
Transcrap Nine ~'p ~X~~
~f~~~~~
' :,~ ' a nucleic acid sequence comprising a sequence in the table below:
~Se '~ dam ~ ~° ~ y;
a , ~ " ~ .r ~. . ~' 'y M78530 PEA 1 node 0 M78530 PEA 1 node 15 M78530 PEA 1 node 16 M78530 PEA-1 node 19 node node node node node node node node node node node node node node node node M78530PEA_1 34 node According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below andlor:
ranscName ~'~~
r '__ a nucleic acid sequence comprising a sequence in the table below:
Segment Name '' ~ ~.~
,: ~ ;:
_".. w_...__~ _. ~ . _.
T48119 node 0 T48119 node I1 T48119 node 13 T48119 node 38 T48119 node 41 T48119 node 45 T48119 node 47 T48119 node 4 T48119 node 8 T48119 node 15 T48119 node 17 T48119 node 20 T48119 node 22 T48119 node 26 T48119 node 28 T48119 node 31 T48119 node 32 T48119 node 33 T48119 node 44 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~~Pro "~am~ ~ ~ ~ °- ~ ~ ~' ~~ ~ ~ ,err.
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~TransCriptName~
~~
-~: ~~,~~' %.~. _ ~ ~:
_ .~n~ t $
I
I
I
I
~HSMUC 1 A T47 PEA_ 1 a nucleic acid sequence comprising a sequence in the table below:
Se~gniien~~
P~ 3 ~ ;.
x ~''~ ~~
~ m HSMUC 1 A PEA node 0 HSMUC 1 A PEA node 14 HSMUC 1 A PEA node 24 HSMUC I A PEA node 29 HSMUC1A PEA node 35 HSMUCIA PEA node 38 HSMUC1A PEA_1 node 3 A PEA node PEA node PEA node PEA node node node HSMUC1A PEA l 20 node node node node node node PEA node node node According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~~a~t~~~e "',a. - 't', ~,, d ~s<r, -e..-..e. .,'~.
a, ~M '-'~'3,~e' a ;. ~~g~~Z
,r~ ", ~
~~s.~, ~~ - ~ "~, m ~'.n~' ~;.z,a.R,~
'~._~ _._.'~_a..m,.
>~a PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
~SMUC 1 A 1 P52 PEA-f-1SMUC1APEA I P53 I A
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSMUCIA PEA-1 P63, comprising a first amino acid sequence being at least 90 % homologous to MTPGTQSPFFLLLLLTVLTVVTGSGHASSTPGGEKETSATQRSSV corresponding to amino acids I - 45 of MUC1 HUMAN, which also corresponds to amino acids 1 - 45 of HSMUCIA PEA-1 P63, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence EEEVSADQVSVGASGVLGSFKEARNAPSFLSWSFSMGPSK corresponding to amino acids 46 - 85 of HSMUC I A PEA-1 P63, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSMUCIA PEA_1 P63, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence EEEVSADQVSVGASGVLGSFKEARNAPSFLSWSFSMGPSK in HSMUCIA PEA 1 P63.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA 1 P2, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGWLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVWVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDN
RYIANTVELRVKISTEVGITNVDLSTVDKDQSIAPKTTRVTYPAKAKGTFIADSHQNFAL
FFQLV DVNTGAELTPHQTFVRLHNQKTGQEV VFVAEPDNKNVYKFELDTSERKlEFDS
ASGTYTLYLI1GDATLKNPILWNV corresponding to amino acids 1 - 498 of RIB2 HUMAN, which also corresponds to amino acids 1 - 498 of T46984 PEA-1 P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VCA corresponding to amino acids 499 - 501 of T46984 PEA_1 P2, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P3, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPV V VVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTP VGDVFELNFMNVKFSSGYYDFLVEVEGDN
RYIANTVELRVKISTEVGITNVDLSTVDKDQSIAPKTTRVTYPAKAKGTFIADSHQNFAL
FFQLVDVNTGAELTPHQ corresponding to amino acids 1 - 433 of RIB2 HUMAN, which also corresponds to amino acids 1 - 433 of T46984 PEA-1 P3, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence ICHIWKLIFLP corresponding to amino acids 434 - 444 of T46984 PEA-1 P3, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA 1 P3, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence ICHIWKLIFLP in T46984 PEA 1 P3.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA_1 P10, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKI-IDVERLKASLDRPFTNLESAFYSIVGLSSL
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDN
FFQLVDVNTGAELTPHQTFVRLHNQKTGQEVVFVAEPDNKNVYKFELDTSERKIEFDS
ASGTYTLYLIIGDATLKNPILWNV corresponding to amino acids 1 - 498 of RIB2 HUMAN, which also corresponds to amino acids 1 - 498 of T46984 PEA 1 P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more 15 preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence LMDQK corresponding to amino acids 499 - 503 of T46984 PEA_1 P10, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an 20 isolated polypeptide encoding for a tail of T46984 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homolo gous to the sequence LMDQK
in T46984 PEA 1 P10.
According to preferred embodiments of the present invention, there is provided an 25 isolated chimeric polypeptide encoding for T46984 PEA_1 P11, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDN
RYIANTVELRVKISTEVGITN V DLSTVDKDQSIAPKTTRVTYPAKAKGTFIADSHQNFAL
FFQLVDVNTGAELTPHQTFV RLHINQKTGQEVVFVAEPDNKNVYKFELDTSERKIEFDS
ASGTYTLYLIIGDATLKNPILWNVADV VIKFPEEEAPSTVLSQNLFTPKQEIQHLFREPEK
RPPTVVSNTFTALILSPLLLLFALWIRIGANVSNFTFAPSTIIFHLGHAAMLGLMYVYWT
QLNMFQTLKYLAILGSVTFLAGNRMLAQQAVKR corresponding to amino acids 1 - 628 of RIB2 HUMAN, which also corresponds to amino acids 1 - 628 of T46984 PEA 1 PI
1.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984_PEA_1-P12, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAF YSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVGDVFELNFMN corresponding to amino acids 1 - 338 of RIB2_HUMAN, which also corresponds to amino acids 1 - 338 of T46984 PEA_1 P12, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence SQDLH corresponding to amino acids 339 -343 of T46984 PEA_1 P12, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA-1 P12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SQDLH in T46984 PEA 1 P 12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA_1 P21, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence M
corresponding to amino acids 1 - I of T46984 PEA-l P21, and a second amino acid sequence being at least 90 % homologous to KACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSSVTQIYHAV
AALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSIVEEIEDLVA
RLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNAIFSKKNFES
LSEAFSVASAAAVLSHNRYH V PV V V VPEGSASDTHEQAILRLQVTN VLSQPLTQATVKL
EHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDNRYIANTVEL
RVKISTEVGITNVDLSTVDKDQSIAPKTTRVTYPAKAKGTFIADSHQNFALFFQLVDVNT
GAELTPHQTFVRLHNQKTGQEVVFVAEPDNKNVYKFELDTSERKIEFDSASGTYTLYLII
GDATLKNPILWNVADVVIKFPEEEAPSTVLSQNLFTPKQEIQHLFREPEKRPPTVVSNTF
TALILSPLLLLFALWIRIGANVSNFTFAPSTIIFHLGHAAMLGLMYVYWTQLNMFQTLKY
LAILGSVTFLAGNRMLAQQAVKRTAH corresponding to amino acids 70 - 631 of RIB2 HUMAN, which also corresponds to amino acids 2 - 563 of T46984 PEA-1 P21, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA 1 P27, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKL,EHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDN
RYIANTVELRVKISTEVGITNVDLSTVDKDQSIAPKTTRVTYPAKAKGTFIADSHQNFA
corresponding to amino acids 1 - 415 of RIB2 HUMAN, which also corresponds to amino acids 1 - 415 of T46984 PEA-1 P27, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence FGSGLVPMSPTSLLLLARLYFTWDMLLCWDSCMSTGLSSTCSRP corresponding to amino acids 416 - 459 of T46984 PEA_1 P27, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA_1 P27, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence FGSGLVPMSPTSLLLLARLYFTWDMLLCWDSCMSTGLSSTCSRP in T46984 PEA 1 P27.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P32, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKI-IDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDN
RYIANTVE corresponding to amino acids 1 - 364 of RIB2 HUMAN, which also corresponds to amino acids 1 - 364 of T46984 PEA_1 P32, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GQVRWLTPVIPALWEAKAGGSPEVRSSILAWPT corresponding to amino acids 365 - 397 of T46984 PEA_1 P32, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA-1 P32, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GQVRWLTPVIPALWEAKAGGSPEVRSSILAWPT in T46984 PEA 1 P32.
According to preferred embodiments of the preset invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P34, comprising a first amino acid sequence being at least 90'% homologous to VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVG corresponding to amino acids 1 - 329 of RIB2 HUMAN, which also corresponds to amino acids 1 - 329 of T46984 PEA 1 P34.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P35, compris ing a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAI corresponding to amino acids 1 - 287 of RIB2 HUMAN, which also corresponds to amino acids 1 -287 of T46984 PEA-1 P35, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 9S%
homologous to a polypeptide having the sequence GCWPSRQSREQHISSRRKMEILKTECQEKESRTIHSMRRKMEKKNFI corresponding to amino acids 288 - 334 of T46984 PEA_1 P35, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA-1 P35, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GCWPSRQSREQHISSRRKMEILKTECQEKESRTIHSMRRKMEKKNFI in T46984 PEA 1 P35.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T469$4 PEA-1 P38, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKI-iDVERLKASLDRPFTNLESAFYSI VGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEAL corresponding to amino acids 1 - 145 of 5 R1B2 HUMAN, which also corresponds to amino acids 1 - 145 of T46984 PEA-1 P38, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MDPDWCQCLQLHFCS corresponding to amino acids 146 - 160 of T46984 PEA-1 P38; wherein said first amino acid sequence and second amino acid sequence 10 are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA_1 P38, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence 15 MDPDWCQCLQLHFCS in T46984 PEA 1 P38.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P39, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLA corresponding to amino acids 1 160 of RIB2 HUMAN, which also corresponds to amino acids 1 - 160 of T46984 PEA
1 P39.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P45, comprising a first amino acid 25 sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCE corresponding to amino acids 1 - 101 of RIB2 HUMAN, which also corresponds to amino acids 1 - 101 of T46984 PEA-1 P45, and a second amino acid sequence being at least 70%, optionally at least 30 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence NSPGSADSIPPVPAG corresponding to amino acids 102 - 1 16 of T46984 PEA_I P45, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984~PEA-1 P45, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NSPGSADSIP.PVPAG in T46984 PEA 1 P45.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P46, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLT.KHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAK corresponding to amino acids 1 - 69 of RIB2,HUMAN, which also corresponds to amino acids 1 - 69 of T46984 PEA I P46, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NSPGSADSIPPVPAG corresponding to amino acids 70 - 84 of T46984~PEA-1 P46, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA-1 P46, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NSPGSADSIPPVPAG in T46984 PEA 1 P46.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA-1 P15, comprising a first amino acid sequence being at least 90 % homologous to MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEE
ETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKL
CEQDSTFDGVTDKPILDCCACGTAKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSH
SKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLN
DAGTDSGVTYESPNKPTIPQEKIRPLTSLDI-IPQSPFYDPEGGSITQVARVVIERIARKGEQ
CNIVPDNVDDIVADLAPEEKDEDDTPETCIYSNWSPWSACSSSTCDKGKRMRQRMLKA
QLDLSVPCPDTQDFQPCMGPGCSDEDGSTCTMSEWITWSPCSISCGMGMRSRERYVKQ
FPEDGSVCTLPTEE corresponding to amino acids 1 - 544 of Q9HCB6, which also corresponds to amino acids 1 - 544 of M78530 PEA_1 P15, a bridging amino acid T
corresponding to amino acid 545 of M78530 PEA-1 P15, a second amino acid sequence being at least 90 % homologous to EKCTVNEECSPSSCLMTEWGEWDECSATCGMGMKKRHRMIKMNPADGSMCKAETSQ
AEKCMMPECHTIPCLLSPWSEWSDCSVTCGKGMRTRQRMLKSLAELGDCNEDLEQVE
KCMLPEC corresponding to amino acids 546 - 665 of Q9HCB6, which also corresponds to amino acids 546 - 665 of M78530 PEA_1 P15, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence RKSWSSSRPITSMFLSPGSPEPASANTARS corresponding to amino acids 666 - 695 of M78530 PEA-1 P15, wherein said first amino acid sequence, bridging amino acid, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M78530 PEA-1 P15, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence RKSWSSSRPITSMFLSPGSPEPASANTARS in M78530 PEA 1 P15.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA_1 P15, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLS corresponding to amino acids 1 - 83 of M78530 PEA_1 P15, a second amino acid sequence being at least 90 % homologous to AAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEEETQFMSNCPVAVTESTPRRRTRIQ
VFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKLCEQDSTFDGVTDKPILDCCACGT
AKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSHSKNYVLWEYGGYASEGVKQVAE
LGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLNVRAAPSAEFSVDRTRHLMSFLTM
MGPSPDWNVGLSAEDLCTKECGW VQKVVQDLIPWDAGTDSGVTYESPNKPTIPQEKIR
PLTSLDHPQSPFYDPEGGSITQVARVVIERIARKGEQCNIVPDNVDDIVADLAPEEKDED
DTPETCIYSNWSPWSACSSSTCDKGKRMRQRMLKAQLDLSVPCPDTQDFQPCMGPGCS
DEDGSTCTMSEWITWSPCSISCGMGMRSRERYVKQFPEDGSVCTLPTEETEKCTVNEEC
SPSSCLMTEWGEWDECSATCGMGMKKRHRM IKMNPADGSMCKAETSQAEKCMMPE
CHTIPCLLSPWSEWSDCSVTCGKGMRTRQRMLKSLAELGDCNEDLEQVEKCMLPEC
corresponding to amino acids 1 - 582 of 094862, which also corresponds to amino acids 84 -665 ofM78530 PEA-1 P15, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence RKSWSSSRPITSMFLSPGSPEPASANTARS corresponding to amino acids 666 - 695 of M78530 PEA-1 P15, wherein said first amino acid sequence, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of M78530 PEA_1 P15, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLS of M78530 PEA 1 P15.
An isolated polypeptide encoding for a tail of M78530 PEA_1 P15, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence RKSWSSSRPITSMFLSPGSPEPASANTARS in M78530 PEA 1 P15.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA-1 P16, comprising a first amino acid sequence being at least 90 % homologous to MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEE
ETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKT.
CEQDSTFDGVTDKPILDCCACGTAKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSH
SKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLN
V corresponding to amino acids 1 - 297 of Q8NCD7, which also corresponds to amino acids 1 -297 of M78530 PEA 1 P16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA-1 P16, comprising a first amino acid sequence being at least 90 % homologous to MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDK VPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEE
ETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTK1CT.
CEQDSTFDGVTDKPILDCCACGTAKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSH
SKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLN
V corresponding to amino acids 1 - 297 of Q9HCB6, which also corresponds to amino acids 1 -297 of M78530 PEA 1 P 16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA_1 P16, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLS corresponding to amino acids 1 - 83 of M78530 PEA 1 P16, and a second amino acid sequence being at least 90 %
homologous to AAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEEETQFMSNCPVAVTESTPRRRTRIQ
VFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKLCEQDSTFDGVTDKPILDCCACGT
AKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSHSKNYVLWEYGGYASEGVKQVAE
LGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLNV corresponding to amino acids 1 -214 of 094862, which also corresponds to amino acids 84 - 297 of M78530 PEA-1 P16, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of M78530 PEA 1 P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLS of M78530 PEA 1 P 16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA_1 P17, comprising a first amino acid sequence being at least 90 % homologous to MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEE
ETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKL
SKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQ corresponding to amino acids I -275 of Q8NCD7, which also corresponds to amino acids I - 275 of M78530 PEA_1 P17, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide 15 having the sequence VRQKNHRMTK corresponding to amino acids 276 - 285 of M78530 PEA-1 P 17, wherein said first amino acid sequence and second amino acid sequence are cont iguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M78530 PEA-1 P17, comprising a polypeptide 20 being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRQKNHRMTK in M78530 PEA 1 P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA-1 P17, comprising a first amino acid 25 sequence being at least 90 % homologous to MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEE
ETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKT.
CEQDSTFDGVTDKPILDCCACGTAKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSH
30 SKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQ corresponding to amino acids 1 -275 of Q9HCB6, which also corresponds to amino acids 1 - 275 of M78530 PEA_1 P17, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRQKNHRMTK corresponding to amino acids 276 - 285 of M78530 PEA,I P17, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M78530 PEA-1 P17, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRQKNHRMTK in M78530 PEA 1 P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA-1 P17, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLS corresponding to amino acids 1 - 83 of M78530 PEA_1 P17, a second amino acid sequence being at least 90 % homologous to AAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEEETQFMSNCPVAVTESTPRRRTRIQ
VFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKLCEQDSTFDGVTDKPILDCCACGT
AKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSHSKNYVLWEYGGYASEGVKQVAE
LGSPVKMEEEIRQQ corresponding to amino acids 1 - 192 of 094862, which also corresponds to amino acids 84 - 275 of M78530 PEA_1 P17, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and rr~st preferably at least 95% homologous to a polypeptide having the sequence VRQKNHRMTK
corresponding to amino acids 276 - 285 of M78530 PEA-1 P17, wherein said first amino acid sequence, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of M78530 PEA-1 P17, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the .seguej~ce MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYTEFSLR
VEGDPDFYKPGTSYRVTLS of M78530 PEA I P17.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M78530 PEA-1 P17, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRQKNHRMTK in M78530 PEA 1 P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T48119 P2, comprising a first amino acid sequence being at least 90 % homologous to MTRQMASSGASGGKIDNSVLVLIVGLSTVGAGAYAYKTMKEDEKRYNERISGLGLTPE
ELPYMRPPLSKELWFSDDPNVTKTLRFKQWNGKERSIYFQPPSFYVSAQDLPHIENGGV
AVLTGKKVVQLDVRDNMVKLNDGSQITYEKCLIATGGTPRSLSAIDRAGAEVKSRTTL
FRKIGDFRSLEKISREVKSITIIGGGFLGSELACALGRKARALGTEVIQLFPEKGNMGKILP
EYLSNWTMEKVRREGVKVMPNAIVQSVGVSSGKLLIKLKDGRKVETDHIVAAVGLEP
NVELAKTGGLEIDSDFGGFRVNAELQARSNIWVAGDAACFYDIKLGRRRVEHHDHAV
VSGRLAGENMTGAAKPYWHQSMFWSDLGPDVGYEAIGLVDSSLPTVGVFAKATAQD
NPKSATEQSGTGIRSESETESEASEITIPPSTPAVPQAPVQGEDYGKGVIFYLRDKVVVGI
VLWNIFNRMPIARKIIKDGEQHEDLNEVAKLFNIHED corresponding to amino acids 50 -613 of PCD8 HI1MAN, which also corresponds to amino acids 1 - 564 of T48119 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T48119 P2, comprising a first amino acid sequence being at least 90 % homologous to MTRQMASSGASGGKIDNSVLVLIVGLSTVGAGAYAYKTMKEDEKRYNERISGLGLTPE
QKQKKAALSASEGEEVPQDKAPSHVPFLLIGGGTAAFAAARSIRARDPGARVLIVSEDP
ELPYMRPPLSKELWFSDDPNVTKTLRFKQWNGKERSIYFQPPSFYVSAQDLPHIENGGV
AVLTGKKVVQLDVRDNMVKLNDGSQITYEKCLIATGGTPRSLSAIDRAGAEVKSRTTL
FRKIGDFRSLEKISREVKSITIIGGGFLGSELACALGRKARALGTEVIQLFPEKGNMGKILP
EYLSNWTMEKVRREGVKVMPNAIVQSVGVSSGKLLIKLKDGRKVETDHIVAAVGLEP
VSGRLAGENMTGAAKPYWHQSMFWSDLGPDVGYEAIGLVDSSLPTVGVFAKATAQD
NPKSATEQSGTGIRSESETESEASEITIPPSTPAVPQAPVQGEDYGKGVIFYLRDKVVVGI
VLWNIFNRMPIARKI.IKDGEQHEDLNEVAKLFNIHED corresponding to amino acids 50 -613 of PCD8~HUMAN, which also corresponds to amino acids 1 - 564 of T48119 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971 P6, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAW VALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
IO KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
LKPEEEAPAPEVGASKPEGIDSRPETLI-IPGRPQPPAEEELCSGKPFDAFTDLKNGSLFAFR
GQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAFTRINCQGKTYLFKGSQYWRFEDGV
LDPDYPRNISDGFDGIPDNVDAALALPAHSYSGRERVYFFKG corresponding to amino acids 1 - 276 of VTNC HUMAN, which also corresponds to amino acids 1 - 276 of 15 T39971 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence TQGVVGD corresponding to amino acids 277 - 283 of T39971 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
20 According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T39971 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TQGWGD
in T39971 P6.
25 According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971 P9, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAWALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
GQYCYELDEKAVRPGYPKLIRDWGIEGPIDAAFTRINCQGKTYLFKGSQYWRFEDGV
LDPDYPRNISDGFDGIPDNVDAALALPAHSYSGRERVYFFKGKQYWEYQFQHQPSQEE
CEGSSLSAVFEI-IFAMMQRDSWEDIFELLFWGRT corresponding to amino acids 1 - 325 of VTNC_HUMAN, which also corresponds to amino acids 1 - 325 of T39971 P9, and a second amino acid sequence being at least 90 % homologous to SGMAPRPSLAKKQRFRI-IRNRKGYRSQRGHSRGRNQNSRRPSRATWLSLFSSEESNLGA
NNYDDYRMDWLVPATCEPIQS VFFFSGDKYYRVNLRTRRV DTV DPPYPRSIAQY W LGC
PAPGHL corresponding to amino acids 357 - 478 of VTNC HUMAN, which also corresponds to amino acids 326 - 447 of T39971 P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T39971 P9, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise TS, having a structure as follows: a sequence starting from any of amino acid numbers 325-x to 325; and ending at any of amino acid numbers 326 + ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971_PI 1, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAWVALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
LKPEEEAPAPEVGASKPEGIDSRPETLHPGRPQPPAEEELCSGKPFDAFTDLKNGSLFAFR
GQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAFTRINCQGKTYLFKGSQYWRFEDGV
LDPDYPRNISDGFDGIPDNVDAALALPAHSYSGRERVYFFKGKQYWEYQFQHQPSQEE
CEGSSLSAVFEHFAMMQRDSWEDIFELLFWGRTS corresponding to amino acids 1 - 326 of VTNC_HUMAN, which also corresponds to amino acids 1 - 326 ofT39971 P11, and a second amino acid sequence being at least 90 % homologous to DKYYRVNLRTRRVDTVDPPYPRSIAQYWLGCPAPGHL corresponding to amino acids 442 - 478 of VTNC_HUMAN, which also corresponds to amino acids 327 - 363 of T39971 P1 I, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T39971 P 11, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more 5 preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise SD, having a structure as follows: a sequence starting from any of amino acid numbers 326-x to 326; and ending at any of amino acid numbers 327 + ((rr2) - x), in which x varies from 0 to rr2.
10 According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971 Pl I, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAWVALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
I S LKPEEEAPAPEVGASKPEGIDSRPETLHPGRPQPPAEEELCSGKPFDAFTDLKNGSLFAFR
GQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAFTRINCQGKTYLFKGSQYWRFEDGV
LDPDYPRNISDGFDGIPDNVDAALALPAHSYSGRERVYFFKGKQYWEYQFQHQPSQEE
CEGSSLSAVFEHFAMMQRDSWEDIFELLFWGRTS corresponding to amino acids 1 - 326 of Q9BSH7, which also corresponds to amino acids 1 - 326 of T39971 PI 1, and a second amino 20 acid sequence being at least 90 % homologous to DKYYRVNLRTRRVDTVDPPYPRSIAQYWLGCPAPGHL corresponding to amino acids 442 - 478 of Q9BSH7, which also corresponds to amino acids 327 - 363 of T39971 P11, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an 25 isolated chimeric polypeptide encoding for an edge portion of T39971 P11, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino . acids in length, wherein at least two amino acids comprise SD, having a structure as follows: a 30 sequence starting from any of amino acid numbers 326-x to 326; and ending at any of amino acid numbers 327 + ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971 P12, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAWVALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
LKPEEEAPAPEVGASKPEGIDSRPETLHPGRPQPPAEEELCSGKPFDAFTDLKNGSLFAFR
GQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAFTRINCQGKTYLFK corresponding to amino acids 1 - 223 of VTNC HUMAN, which also corresponds to amino acids 1 -223 of T39971 P12, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence VPGAVGQGRKHLGRV corresponding to amino acids 224 - 238 of T39971 P12, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T39971 P12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VPGAVGQGRKHLGRV in T39971 P12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971 P12, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAWVALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
LKPEEEAPAPEVGASKPEGIDSRPETLHPGRPQPPAEEELCSGKPFDAFTDLKNGSLFAFR
GQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAFTRINCQGKTYLFK corresponding to amino acids 1 - 223 of Q9BSH7, which also corresponds to amino acids 1 - 223 of T39971 P12, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VPGAVGQGRKHLGRV corresponding to amino acids 238 of T39971 P12, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T39971 P12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VPGAVGQGRKHLGRV in T39971 P12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 244808 PEA-1 P5, comprising a first amino acid -sequence being at least 90 % homologous to MLLPQLCWLPLLAGLLPPV.PAQKFSALTFLRVDQDKDKDCSLDCAGSPQKPLCASDGR
TFLSRCEFQRAKCKDPQLEIAYRGNCKDVSRCVAERKYTQEQARKEFQQVFIPECNDD
GTYSQVQCHSYTGYCWCVTPNGRPISGTAVAHKTPRCPGSVNEKLPQREGTGKTDDAA
APALETQPQGDEEDIASRYPTLWTEQVKSRQNKTNKNSVSSCDQEHQSALEEAKQPKN
DNWIPECAHGGLYKPVQCHPSTGYCWCVLVDTGRPIPGTSTRYEQPKCDNTARAHPA
KARDLYKGRQLQGCPGAKKHEFLTSVLDALSTDMVHAASDPSSSSGRLSEPDPSHTLEE
RVVHWYFKLLDKNSSGDIGKKEIKPFKRFLRKKSKPKKCVKKFVEYCDVNNDKSISVQ
ELMGCLGVAKEDGKADTKKRHTPRGHAESTSNRQ corresponding to amino acids 1 - 441 of SM02 HUMAN, which also corresponds to amino acids 1 - 441 of 244808 PEA-1 PS, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DAMVVSSRPKATTHRKSRTLSRR corresponding to amino acids 442 - 464 of 244808 PEA-1 P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 244808 PEA_1 P5, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DAMVVSSRPKATTHRKSRTLSRR in 244808 PEA 1 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 244808 PEA-1 P6, comprising a first amino acid sequence being at least 90 % homologous to MLLPQLCWLPLLAGLLPPVPAQKFSALTFLRVDQDKDKDCSLDCAGSPQKPLCASDGR
TFLSRCEFQRAKCKDPQLEIAYRGNCKDVSRCVAERKYTQEQARKEFQQVFIPECNDD
GTYSQVQCHSYTGYCWCVTPNGRPISGTAVAHKTPRCPGS VNEKLPQREGTGKTDDAA
APALETQPQGDEEDIASRYPTLWTEQVKSRQNKTNKNSVSSCDQEHQSALEEAKQPKN
DNVVIPECAHGGLYKPVQCI-IPSTGYCWCVLVDTGRPIPGTSTRYEQPKCDNTARAHPA
KARDLYKGRQLQGCPGAKKHEFLTSVLDALSTDMVHAASDPSSSSGRLSEPDPSHTLEE
RV VH WYFKLLDKNSSGDIGKKEIKPFKRFLRKKSKPKKCVKKFV EYCD VNNDKSISVQ
ELMGCLGVAKEDGKADTKKRH corresponding to amino acids 1 - 428 of SM02_HUMAN, which also corresponds to amino acids 1 - 428 of Z44808,PEA 1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence RSKRNL corresponding to amino acids 429 - 434 of 244808 PEA-1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 244808 PEA_I P6, comprising a polypeptide being I S at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence RSKRNL
in 244808 PEA I P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 244808 PEA-1 P7, comprising a first amino acid sequence being at least 90 % homologous to MLLPQLCWLPLLAGLLPPVPAQKFSALTFLRVDQDKDKDCSLDCAGSPQKPLCASDGR
TFLSRCEFQRAKCKDPQLEIAYRGNCKDVSRCVAERKYTQEQARKEFQQVFIPECNDD
GTYSQVQCHSYTGYCWCVTPNGRPISGTAVAHKTPRCPGSVNEKLPQREGTGKTDDAA
APALETQPQGDEEDIASRYPTLWTEQVKSRQNKTNKNSVSSCDQEHQSALEEAKQPKN
DNVVIPECAHGGLYKPVQCHPSTGYCWCVLVDTGRPIPGTSTRYEQPKCDNTARAHPA
KARDLYKGRQLQGCPGAKKHEFLTSVLDALSTDMVHAASDPSSSSGRLSEPDPSHTLEE
RVVHWYFKLLDKNSSGDIGKKEIKPFKRFLRKKSKPKKCVKKFVEYCDVNNDKSISVQ
ELMGCLGVAKEDGKADTKKRHTPRGHAESTSNRQ corresponding to amino acids 1 - 441 of SM02 HUMAN, which also corresponds to amino acids 1 - 441 of 244808 PEA 1 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a '7 9 polypeptide having the sequence LLWLRGKVSFYCF corresponding to amino acids 442 of 244808 PEA-1 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 244808 PEA_1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence LLWLRGKVSFYCF in 244808 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 244808 PEA_1 P11, comprising a first amino acid sequence being at least 90 % homologous to MLLPQLCWLPLLAGLLPPVPAQKFSALTFLRVDQDKDKDCSLDCAGSPQKPLCASDGR
TFLSRCEFQRAKCKDPQLEIAYRGNCKDVSRCVAERKYTQEQARKEFQQVFIPECNDD
GTYSQVQCHSYTGYCWCVTPNGRPISGTAVAHKTPRCPGSVNEKLPQREGTGKT
1 S corresponding to amino acids 1 - 170 of SM02 HUMAN, which also corresponds to amino acids 1 - 170 of 244808 PEA_1 P11, and a second amino acid sequence being at least 90 homologous to DIASRYPTLWTEQVKSRQNKTNKNSVSSCDQEHQSALEEAKQPKNDNV VIPECAHGGL
YKPVQCHPSTGYCWCVLVDTGRPIPGTSTRYEQPKCDNTARAHPAKARDLYKGRQLQ
GCPGAKKHEFLTSVLDALSTDMVHAASDPSSSSGRLSEPDPSHTLEERVVHWYFKLLD
KNSSGDIGKKEIKPFKRFLRKKSKPKKCVKKFVEYCDVNNDKSISVQELMGCLGVAKE
DGKADTKKRHTPRGHAESTSNRQPRKQG corresponding to amino acids 188 - 446 of SM02 HUMAN, which also corresponds to amino acids 171 - 429 of 244808 PEA,1 P1 l, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of 244808 PEA-1 P11, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise TD, having a structure as follows: a sequence starting from any of amino acid numbers 170-x to -170; and ending at any of amino acid numbers 171+ ((,r2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for S67314 PEA_I P4, comprising a first amino acid 5 sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence M VDAFLGTWKLV DSKNFDDYMKSLGVGFATRQVASMTKPTTIIEKNGDILTLKTHSTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of FABH HUMAN, which also corresponds to amino 10 acids 1 - 116 of 567314 PEA-1 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence VRWATLELYLIGYYYCSFSQACSKKPSPPLRAVEAGTREWLWVRVVSGGNFLCSGFGL
TQAGTQILPYRLHDCGQITFSKCNCKTGINNTNLVGLLGSL corresponding to amino acids 15 117 - 215 of 567314 PEA_1 P4, wherein said firstand second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA-1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at 20 least about 90% and most preferably at least about 95% homologous to the sequence VRWATLELYLIGYYYCSFSQACSKKPSPPLRAVEAGTREWLWVRVVSGGNFLCSGFGL
TQAGTQILPYRLHDCGQITFSKCNCKTGINNTNLVGLLGSL in 567314 PEA 1 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 567314 PEA_1 P4, comprising a first amino acid 25 sequence being at least 90 % homologous to MVDAFLGTWKLVDSKNFDDYMKSLGVGFATRQVASMTKPTTIIEKNGDILTLKTHSTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of AAP35373, which also corresponds to amino acids 1 -116 of 567314 PEA-1 P4, and a second amino acid sequence being at least 70%, optionally at 30 least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRWATLELYLIGYYYCSFSQACSKKPSPPLRAVEAGTREWLWVRVVSGGNFLCSGFGL
TQAGTQIL.PYRLHDCGQITFSKCNCKTGINNTNLVGLLGSL corresponding to amino acids 117 - 215 of 567314 PEA_1 P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA_1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRWATLELYLIGYYYCSFSQACSKKPSPPLRAVEAGTREWLWVRVVSGGNFLCSGFGL
TQAGTQILPYRLHDCGQITFSKCNCKTGINNTNLVGLLGSL in 567314 PEA_I P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 567314 PEA_1 P5, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MVDAFLGTWKLVDSKNFDDYMKSLGVGFATRQVASMTKPTTIIEKNGDILTLKTHSTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of FABH HUMAN, which also corresponds to amino acids 1 - 116 of S67314 PEA_1 P5, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DVLTAWPSIYRRQVKVLREDEITILPWHLQWSREKATKLLRPTLPSYNNHGWEELRVG
KSIV corresponding to amino acids 117 - 178 of S67314 PEA_1 P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA_1 P5, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DVLTAWPSIYRRQVKVLREDEITILPWHLQWSREKATKLLRPTLPSYNNHGWEELRVG
KSN in 567314 PEA 1 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 567314 PEA-1 P5, comprising a first amino acid' sequence being at least 90 % homologous to MVDAFLGTWKLVDSKNFDDYMKSLGVGFATRQVASMTKPTTI f EKNGDILTLKTI-ISTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of AAP35373, which also corresponds to amino acids 1 116 of 567314 PEA-1 P5, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DVLTAWPSIYRRQVKV LREDEITILPWHLQWSREKATKLLRPTLPSYNNHGWEELRVG
KSIV corresponding to amino acids 117 - 178 of S67314 PEA-1 PS, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA-1 PS, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DVLTAWPSIYRRQVKVLREDEITILPWHLQWSREKATKLLRPTLPSYNNHGWEELRVG
KSIV in S67314 PEA 1 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimerie polypeptide encoding for S67314 PEA-1 P6, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MVDAFLGTWKLVDSKNFDDYMKSLGVGFATRQVASMTKPTTIIEKNGDILTLKTHSTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of FABH HUMAN, which also corresponds to amino acids 1 - 116 of S67314 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence MEKLQLRNVK
corresponding to amino acids 117 - 126 of 567314 PEA_1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MEKLQLRNVK in S67314 PEA-1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 567314 PEA-1 P6, comprising a first amino acid sequence being at least 90 % homologous to MVDAFLGTWKLVDSKNFDDYM.KSLGVGFATRQVASMTKPTTIIEKNGDILTLKTHSTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of AAP35373, which also corresponds to amino acids 1 -116 of S67314 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MEKLQLRNVK corresponding to amino acids 117 - 126 of 567314 PEA-1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MEKLQLRNVK in S67314 PEA 1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 567314 PEA-1 P7, comprising a first amino acid sequence being at least 90 % homologous to MVDAFLGTWKLVDSKNFDDYMKSL
corresponding to amino acids 1 - 24 of FABH HUMAN, which also corresponds to amino acids 1 - 24 of 567314 PEA_l P7, second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence AHILITFPLPS corresponding to amino acids 25 - 35 of 567314 PEA 1 P7, and a third amino acid sequence being at least 90 homologous to GVGFATRQVASMTKPTTIIEKNGDILTLKTHSTFKNTEISFKLGVEFDETTADDRKVKSI
VTLDGGKLVHLQKWDGQETTLVRELIDGKLILTLTHGTAVCTRTYEKEA corresponding to amino acids 25 - 133 of FABH HUMAN, which also corresponds to amino acids 36 - 144 of 567314 PEA-1 P7, wherein said first, second, third and fourth amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of S67314 PEA_1 P7, comprising an amino acid sequence being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence encoding for AHILITFPLPS, corresponding to S67314 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for S67314 PEA-1 P7, corr~rising a first amino acid sequence being at least 90 % homologous to MVDAFLGTWKLVDSKNFDDYMKSL
corresponding to amino acids 1 - 24 of AAP35373, which also corresponds to amino acids 1 -24 of S67314 PEA-1 P7, second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence AHILITFPLPS corresponding to amino acids 25 - 35 of 567314 PEA-I P7, and a third amino acid sequence being at least 90 % homologous to GVGFATRQVASMTKPTTIIEKNGDILTLKTHSTFKNTEISFKLGVEFDETTADDRKVKSI
VTLDGGKLVHLQKWDGQETTLVRELIDGKLILTLTHGTAVCTRTYEKEA corresponding to amino acids 25 - 133 of AAP35373, which also corresponds to amino acids 36 -144 of 567314 PEA-1 P7, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of S67314 PEA 1 P7, comprising an amino acid sequence being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence encoding for AHILITFPLPS, corresponding to 567314 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 239337 PEA 2 PEA-1 P4, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWLPLSGAA corresponding to amino acids 1 - 9 of 239337 PEA 2 PEA-1 P4, and a second amino acid sequence being at least 90 homologous to MKKLMVVLSLIAAAWAEEQNKLVHGGPCDKTSHPYQAALYTSGHLLCGGVLIHPLWV
LTAAHCKKPNLQVFLGKHNLRQRESSQEQSSVVRAVIHPDYDAASHDQDIMLLRLARP
S AKLSELIQPLPLERDCSANTTSCHILGWGKTADGDFPDTIQCAYIHLVSREECEHAYPGQ
YTNWIQKTIQAK corresponding to amino acids 1 - 244 of KLK6 HUMAN, which also corresponds to amino acids 10 - 2S3 of 239337 PEA 2 PEA_1 P4, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
10 According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of 239337 PEA 2 PEA-1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 8S%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence MWLPLSGAA of 239337 PEA 2 PEA-1 P4.
1 S According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 239337 PEA 2 PEA-1 P9, comprising a first amino acid sequence being at least 90 % homologous to MKKLMVVLSLIAAAWAEEQNKLVHGGPCDKTSHPYQAALYTSGHLLCGGVLIHPLWV
LTAAHCKKPNLQVFLGKHNLRQRESSQEQSSVVRAVIHPDYDAASHDQDIMLLRLARP
20 AKLSELIQPLPLERDCSANTTSCHILGWGKTADG corresponding to amino acids 1 - 149 of KLK6_HUMAN, which also corresponds to amino acids 1 - 149 of 239337 PEA 2 PEA-1 P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 8S%, more preferably at least 90% and most preferably at least 9S% homologous to a polypeptide having the sequence Q corresponding to amino acids 1 SO -2S 1 SO of 239337 PEA 2 PEA-1 P9, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P10, comprising a first amino acid sequence being at least 90 % homologous to FYYNISE corresponding to amino acids 1 - 67 of PLTP HUMAN, which also corresponds to amino acids I - 67 of HUMPHOSLIP PEA 2 PIO, and a second amino acid sequence being at least 90 % homologous to KVYDFLSTFITSGMRFLLNQQICPVLYHAGTVLLNSLLDTVPVRS SVDELVGIDYSLMK
DPVASTSNLDMDFRGAFFPLTERNWSLPNRAVEPQLQEEERMVYVAFSEFFFDSAMES
YFRAGALQLLLVGDKVPHDLDMLLRATYFGSIVLLSPAVIDSPLKLELRVLAPPRCTIKP
SGTTISVTASVTIALVPPDQPEVQLSSMTMDARLSAKMALRGKALRTQLDLRRFRIYSN
HSALESLALIPLQAPLKTMLQIGVMPMLNERTWRGVQIPLPEGINFVHEVVTNHAGFLTI
GADLHFAKGLREVIEKNRPADVRASTAPTPSTAAV corresponding to amino acids 163 -493 of PLTP HUMAN, which also corresponds to amino acids 68 - 398 of HUMPHOSLIP PEA 2 P10, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMPHOSLIP PEA 2 P10, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EK, having a structure as follows: a sequence starting from any of amino acid numbers 67-x to 67; and ending at any of amino acid numbers 68+ ((n-2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P12, comprising a first amino acid sequence being at least 90 % homologous to MALFGALFLALLAGAHAEFPGCKIRVTSKALELVKQEGLRFLEQELETITIPDLRGKEGH
FYYNISEVKVTELQLTSSELDFQPQQELMLQITNASLGLRFRRQLLYWFFYDGGYINAS
AEGVSIRTGLELSRDPAGRMKVSNVSCQASVSRMHAAFGGTFKKVYDFLSTFITSGMRF
LLNQQICPVLYHAGTVLLNSLLDTVPVRSSVDELVGIDYSLMKDPVASTSNLDMDFRG
AFFPLTERNWSLPNRAVEPQLQEEERMVYVAFSEFFFDSAMESYFRAGALQLLLVGDK
VPHDLDMLLRATYFGSIVLLSPAVIDSPLKLELRVLAPPRCTIKPSGTTISVTASVTIALVP
PDQPEVQLSSMTMDARLSAKMALRGKALRTQLDLRRFRIYSNHSALESLALIPLQAPLK
TMLQIGVMPMLN corresponding to amino acids 1 - 427 of PLTP HUMAN, which also corresponds to amino acids 1 - 427 of HUMPHOSLIP PEA 2 P12, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85'%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence i, GKAGV corresponding to amino acids 428 - 432 of HUMPI-IOSLIP PEA 2 P 12, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMPHOSLIP PEA,2 P 12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence GKAGV in HUMPHOSLIP~PEA 2 P 12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P31, comprising a first amino acid sequence being at least 90 % homologous to MALFGALFLALLAGAHAEFPGCKIRVTSKALELVKQEGLRFLEQELETITIPDLRGKEGH
I 5 FYYNISE corresponding to amino acids 1 - 67 of PLTP HUMAN, which also corresponds to amino acids 1 - 67 of HUMPHOSLIP PEA 2 P31, and a second amino acid sequence being at .yp: _ least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PGLERGADKFPVVGGSSLFLALDLTLRPPVG corresponding to amino acids 68 - 98 of HUMPHOSLIP PEA 2 P31, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMPHOSLIP PEA 2 P31, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence PGLERGADKFPVVGGSSLFLALDLTLRPPVG in HUMPHOSLIP PEA 2 P31.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P33, comprising a first amino acid sequence being at least 90 % homologous to MALFGALFLALLAGAHAEFPGCKIRVTSKALELVKQEGLRFLEQELETITIPDLRGKEGH
FYYNISEVKVTELQLTSSELDFQPQQELMLQITNAS LGLRFRRQLLYWFFYDGGYINAS
AEGVSIRTGLELSRDPAGRMKVSNVSCQASVSRMHAAFGGTFKKVYDFLSTFITSGMRF
LLNQQ corresponding to amino acids 1 - 183 of PLTP HUMAN, which also corresponds to amino acids 1 - 183 of HUMPHOSLIP PEA 2 P33, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VWAATGRRVARVGMLSL corresponding to amino acids 184 - 200 of HUMPHOSLIP PEA 2 P33, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMPHOSLIP PEA 2 P33, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence VWAATGRRVARVGMLSL in HUMPHOSLIP PEA 2 P33.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P34, comprising a first amino acid sequence being at least 90 % homologous to MALFGALFLALLAGAHAEFPGCKIRVTSKALELVKQEGLRFLEQELETITIPDLRGKEGH
FYYNISEVKVTELQLTSSELDFQPQQELMLQITNASLGLRFRRQLLYWFFYDGGYINAS
AEGVSIRTGLELSRDPAGRMKVSNVSCQASVSRMHAAFGGTFKKVYDFLSTFITSGMRF
LLNQQICPVLYHAGTVLLNSLLDTVPV corresponding to amino acids 1 - 205 of PLTP HUMAN, which also corresponds to amino acids 1 - 205 of HUMPHOSLIP PEA 2 P34, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence LWTSLLALTIPS corresponding to amino acids 206 - 217 of HUMPHOSLIP PEA 2 P34, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMPHOSLIP PEA 2 P34, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence LWTSLLALTIPS in HUMPHOSLIP PEA 2 P34.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P35, comprising a first amino acid sequence being at least 90 % homologous to MALFGALFLALLAGAI IAEFPGCKIRVTSKALELVKQEGLRFLEQELETITIPDLRGKEGH
FYYNISEVKVTELQLTSSELDFQPQQELMLQITNASLGLRFRRQLLYWF corresponding to amino acids I - 109 of PLTP HUMAN, which also corresponds to amino acids 1 -109 of HUMPHOSLIP PEA 2 P35, a second amino acid sequence bridging amino acid sequence comprising of L, a third amino acid sequence being at least 90 % homologous to KVYDFLSTFITSGMRFLLNQQ corresponding to amino acids I 63 - 183 of PLTP HUMAN, which also corresponds to amino acids 111 - 131 of HUMPHOSLIP PEA 2 P35, and a fourth amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VWAATGRRVARVGMLSL corresponding to amino acids 132 - 148 of HUMPHOSLIP PEA 2 P35, wherein said first amino acid sequence, second amino acid sequence, third amino acid sequence and fourth amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of HUMPHOSLIP PEA 2 P35, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise FLK
having a structure as follows (numbering according to HUMPHOSLIP PEA 2 P35): a sequence starting from any of amino acid numbers 109-x to 109; and ending at any of amino acid numbers 111 +
((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMPHOSLIP PEA 2 P35, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence VWAATGRRVARVGMLSL in HUMPHOSLIP PEA 2 P35.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832-P7, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNG corresponding to amino acids 12 - 223 of GILT HUMAN, which also corresponds to amino acids 1 - 212 of T59832 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, 10 more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRIFLALSLTLIVPWSQGWTRQRDQR corresponding to amino acids 213 - 238 of T59832 P7, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an 15 isolated polypeptide encoding for a tail of T59832 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRIFLALSLTLIVPWSQGWTRQRDQR in T59832 P7.
According to preferred embodiments of the present invention, there is provided an 20 isolated chimeric polypeptide encoding for T59832 P9, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
25 ECAMGDRGMQLMHANAQRTDALQPPHE corresponding to amino acids 12 - 214 of GILT HUMAN, which also corresponds to amino acids 1 - 203 of T59832 P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR corresponding to 30 amino acids 204 - 244 of T59832 P9, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80°I°, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR in T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P12, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVE corresponding to amino acids 12 - 141 of GILT HUMAN, which also corresponds to amino acids 1 - 130 of T59832 P12, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGD RGMQLMHANAQRTDALQPPHEYVPW VTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 173 - 261 of GILT HUMAN, which also corresponds to amino acids 131 - 219 of T59832 P12, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P12, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EC, having a structure as follows: a sequence starting from any of amino acid numbers 130-x to 130; and ending at any of amino acid numbers 131+ ((n-2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P18, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYK corresponding to amino acids 12 - 55 of GILT HUMAN, which also corresponds to amino acids I - 44 of T59832 P18, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 173 - 261 of GILT HUMAN, which also corresponds to amino acids 45 - 133 of T59832 P 18, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P18, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise KC, having a structure as follows: a sequence starting from any of amino acid numbers 44-x to 44; and ending at any of amino acid numbers 45+ ((rr2) - x), in which x varies from 0 to rr2.
I 5. According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P4, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVWHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWLFGMGNEVDVH
AAFFHGQALTNKNYRIDTINLFPATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFF
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTWSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSWWL
FSAGNEADVHGIYFSGNTYLWRGERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDH
YTGGMKQKYTVNQCRRQSEDSTFYLGERTWIAAVEVEWDYSPQREWEKELHHLQEQ
NVSNAFLDKGEFYIGSKYKKVVYRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDK
YSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYS
DHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHT
V H FHGHSFQYKHRGVYSSDVFDIFPGTYQTLEMFPRTPGIWLLHCHVTDHIHAGMETT
YTVLQNE corresponding to amino acids 1 - 1060 of CERU HUMAN, which also corresponds to amino acids 1 - 1060 of HSCP2 PEA-1 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GGTSM
corresponding to amino acids 1061 - 1065 of HSCP2 PEA_1 P4, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA_1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GGTSM in HSCP2 PEA 1 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P8, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKL,ISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
AAFFHGQALTNKNYRIDTINLFPATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFF
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSVV WYL
FSAGNEADVHGIYFSGNTYLWRGERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDH
YTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQ
NVSNAFLDKGEFYIGSKYKKVVYRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDK
VKIIFKNMATRPYSIHAHGVQTESSTVTPTLPGETLTYVWKIPERSGAGTEDSACIPWAY
YSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYS
DHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHT
VHFHGHSFQYK corresponding to amino acids 1 - 1006 ofCERU HUMAN, which also corresponds to amino acids 1 - 1006 of HSCP2 PEA-1 P8, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence KCFQEHLEFGYSTAM corresponding to amino acids 1007 - 1021 of HSCP2 PEA-1 P8, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA_1 P8, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KCFQEHLEFGYSTAM in HSCP2 PEA-1 P8.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA_1 P14, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
AAFFHGQALTNKNYRIDTINLFPATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFF
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMH corresponding to amino acids 1 - 621 of CERU HUMAN, which also corresponds to amino acids 1 - 621 of HSCP2_PEA-1 P14, a second amino acid sequence bridging amino acid sequence comprising of W, and a third amino acid sequence being at least 90 % homologous to TFNVECLTTDHYTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQRE
WEKELHHLQEQNVSNAFLDKGEFYIGSKYKKVVYRQYTDSTFRVPVERKAEEEHLGIL
GTEDSACIPWAYYSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENE
SWYLDDNIKTYSDHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMH V GDEVN WYL
MGMGNEIDLHTVHFHGHSFQYKHRGVYSSDVFDIFPGTYQTLEMFPRTPGI W LLHCHV
TDHIHAGMETTYTVLQNEDTKSG corresponding to amino acids 694 - 1065 of 10 CERU HUMAN, which also corresponds to amino acids 623 - 994 of HSCP2 PEA-1 P14, wherein said first amino acid sequence, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of HSCP2 PEA_1 P14, comprising a 15 polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise HWT having a structure as follows (numbering according to HSCP2 PEA_1 P14): a sequence starting from any of amino acid 20 numbers 621-x to 621; and ending at any of amino acid numbers 623 + ((rr2) -x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA 1 P15, comprising a first amino acid sequence being at least 90 % homologous to RIGRLYKKALYLQYTDETFRTTIEKPV WLGFLGPIIKAETGDKVWHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFWMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSVVWYL
FSAGNEADVHGIYFSGNTYLWRGERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDH
YTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQ
NV SNAFLDKGEFYIGSKYKKWYRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDK
VKIIFKNMATRPYSIHAHGVQTESSTVTPTLPGETLTYVWKIPERSGAGTEDSACIPWAY
YSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYS
DHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHT
VHFHGHSFQYKHRGVYSSDVFDIFPGTYQTLEMFPRTPGIWLLHCI-IVTDHIHAGMETT
YTVLQNE corresponding to amino acids I - 1060 of CERU I-IUMAN, which also corresponds to amino acids 1 - 1060 of HSCP2 PEA-1 P15, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and I S most preferably at least 95% homologous to a polypeptide having the sequence GEYPASSETHRRIWNVIYPITVSVIILFQISTKE corresponding to amino acids 1061 - 1094 of HSCP2 PEA_1 P15, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA-1 P 15, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GEYPASSETHRRIWNVIYPITVSVIILFQISTKE in HSCP2 PEA_1 P15.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P2, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFWMFSWDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
AAFFHGQALTNKNYRIDTINLFPATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFF
QV QECN:KSSSKDNIRGKHV RHYYIAAEEIIWNYAPSGIDIFTKEN LTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSI EPIG VRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNI RMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSVVWYL
YTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQ
corresponding to amino acids 1 - 761 of CERU HUMAN, which also corresponds to amino acids 1 - 761 of HSCP2 PEA_1 P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence K
corresponding to amino acids 762 - 762 of HSCP2 PEA_1 P2, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA,1 P16, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSWDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
AAFFHGQALTNKNYRIDTINLFPATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFF
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLWREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSWWYL
FSAGNEADVHGIYFSGNTYLWRGERRDTAN LFPQTSLTLHMWDTEGTFNVECLTTDH
YTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQ
NVSNAFLDKGEFYIGSKYKKVWRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDK
V KI IFKNMATRPYSIHAHG VQTESSTVTPTLPGETLTYV W KIPERSGAGTEDSACIPWA Y
YSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYS
DHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHT
VHFHGHSFQYKH corresponding to amino acids 1 - 1007 of CERU HUMAN, which also corresponds to amino acids 1 - 1007 of HSCP2 PEA_1 P16, and a second amino acid sequence being at least 70%, optiona lly at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence LLRLTGEYGM corresponding to amino acids 1008 - 1017 of HSCP2 PEA_1 P16, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA_I P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence IS LLRLTGEYGM in HSCP2 PEA 1 P16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P6, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSVVWYL
FSAGNEADVHGIYFSGNTYLWRGERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDH
YTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQ
VKIIFKNMATRPYSIHAHGVQTESSTVTPTLPGETLTYVWKIPERSGAGTEDSACIPWAY
YSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYS
DHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHT
VHFHGHSFQYK corresponding to amino acids 1 - 1006 of CERU HUMAN, which also corresponds to amino acids 1 - 1006 of HSCP2 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GSL
corresponding to amino acids 1007 - 1009 of HSCP2 PEA_1 P6, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA_l P22, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHE corresponding to amino acids 1 - 131 of CERU HUMAN, which also corresponds to amino acids 1 - 131 of HSCP2 PEA-1 P22, a second amino acid sequence bridging amino acid sequence comprising of A, and a third amino acid sequence~being at least 90 % homologous to ATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFFQVQECNKSSSKDNIRGKHVRHY
YIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQGTTRIGGSYKKLVYREYTDASFTN
RKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAYPLSIEPIGVRFNKNNEGTYYSPNY
NPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNADPVCLAKMYYSAVDPTKDIFTGLI
GPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENESLLLEDNIRMFTTAPDQVDKEDE
DFQESNKMHSMNGFMYGNQPGLTMCKGDSWWYLFSAGNEADVHGIYFSGNTYLWR
GERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDHYTGGMKQKYTVNQCRRQSEDS
TFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQNVSNAFLDKGEFYIGSKYKKW
YRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDKVKIIFKNMATRPYSIHAHGVQTE
SSTVTPTLPGETLTYVWKIPERSGAGTEDSACIPWAYYSTVDQVKDLYSGLIGPLIVCRR
PYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYSDHPEKVNKDDEEFIESNKMHAI
NGRMFGNLQGLTMI-IVGDEVNWYLMGMGNEIDLHTVHFI-IGHSFQYKHRGVYSSDVF
DIFPGTYQTLEMFPRTPGIWLLHCHVTDHIHAGMETTYTVLQNEDTKSG corresponding to amino acids 262 - 1065 of CERU HUMAN, which also corresponds to amino acids 936 of HSCP2 PEA_1 P22, wherein said first amino acid sequence, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of HSCP2_PEA-1 P22, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EAV having a structure as follows (numbering according to HSCP2 PEA-1 P22): a sequence starting from any of amino acid numbers 131-x to 131; and ending at any of amino acid numbers 133 + ((n-2) -x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA_I P24, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MPLTMGKRNLFLLTP corresponding to amino acids 1 - 15 of HSCP2 PEA_1 P24, and a second amino acid sequence being at least 90 % homologous to VNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVHAAFFHGQALTNKNYRIDTINLFP
ATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFFQVQECNKSSSKDNIRGKHVRHY
YIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQGTTRIGGSYKKLWREYTDASFTN
RKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAYPLSIEPIGVRFNKNNEGTWSPNY
NPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNADPVCLAKMYYSAVDPTKDIFTGLI
GPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENESLLLEDNIRMFTTAPDQVDKEDE
DFQESNKMHSMNGFMYGNQPGLTMCKGDSVVWYLFSAGNEADVHGIYFSGNTYLWR
GERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDHYTGGMKQKYTVNQCRRQSEDS
TFYLGERTWIAAVEVEWDYSPQREWEKELHHLQEQNVSNAFLDKGEFYIGSKYKKW
YRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDKVKIIFKNMATRPYSIHAHGVQTE
SSTVTPTLPGETLTYV WKIPERSGAGTEDSACIPWAYYSTVDQVKDLYSGLIGPLIVCRR
PYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYSDHPEKVNKDDEEFIESNKMHAI
NGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHTVHFI-IGHSFQYKHRGVYSSDVF
DIFPGTYQTLEMFPRTPGIWLLHCHVTDHIHAGMETTYTVLQNEDTKSG corresponding to amino acids 262 - 1065 of CERU HUMAN, which also corresponds to amino acids of HSCP2 PEA-1 P24, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of HSCP2 PEA-1 P24, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MPLTMGKRNLFLLTP of HSCP2 PEA 1 P24.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P25, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPV WLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMH corresponding to amino acids 1 - 621 of CERU HUMAN, which also corresponds to amino acids 1 - 621 of HSCP2 PEA-1 P25, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence CKYCIIHQSTKLF corresponding to amino acids 622 - 634 of HSCP2 PEA-I P25, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA_1 P25, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence CKYCIIHQSTKLF in HSCP2 PEA-1 P25.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P33, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HSHIDAPKDIASGLIGPLIICKK corresponding to amino acids 1 - 202 of CERU HUMAN, which also corresponds to amino acids 1 - 202 of HSCP2 PEA_1 P33, and a second amino acid sequence being at least 70°l°, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GTSSPYCTCYMTKRQGQGSLSFKKKSSLLC corresponding to amino acids 203 - 232 of HSCP2 PEA_1 P33, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA_1 P33, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GTSSPYCTCYMTKRQGQGSLSFKKKSSLLC in HSCP2_PEA-1 P33.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN_PEA_1 P5, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQL.RCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQtaPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EV V VAE VGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
T corresponding to amino acids 1 - 1525 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 1525 of HUMTEN PEA-1 P5, a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence TEPKPQLGTLIFSNITPKSFNMSWTTQAGLFAKIVINVSDAHSLHESQQFTVSGDAKQAH
ITGLVENTGYDVS VAGTTLAGDPTRPLTAFVI corresponding to amino acids 1526 - 1617 of HUMTEN PEA_1 P5, and a third amino acid sequence being at least 90 %
homologous to TEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTWDSGKLLDPQEFTLSGTQRKLE
LRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTADE
GVFDNFVLKIRDTKKQSEPLEITLLAPERTRDLTGLREATEYEIELYGISKGRRSQTVSAI
ATTAMGSPKEV IFSDITENSATV SWRAPTAQVESFRITYV PITGGTPSM VTV DGTKTQTR
LVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIATV
PSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGDK
AQALEVFCDMTSDGGG WIVFLRRKNGRENFYQNWKAYAAGFGDRREEFWLGLDNLN
KITAQGQYELRVDLRDHGETAFAVYDKFSVGDAKTRYKLKVEGYSGTAGDSMAYHN
GRSFSTFDKDTDSAITNCALSYKGAFWYRNCHRVNLMGRYGDNNHSQGVNWFHWKG
HEHSIQFAEMKLRPSNFRNLEGRRKRA corresponding to amino acids 1526 - 2201 of TENA HUMAN V1, which also corresponds to amino acids 1618 - 2293 of HUMTEN PEA_1 P5, wherein said first amino acid sequence, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of HUMTEN PEA_1 P5, comprising an amino acid sequence being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence encoding for TEPKPQLGTLIFSNITPKSFNMSWTTQAGLFAKIVINVSDAHSLHESQQFTVSGDAKQAH
ITGLVENTGYDVSVAGTTLAGDPTRPLTAFVI, corresponding to HUMTEN PEA_1 PS.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_1 P6, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPV SARV ATYLPAPEGLKFKSIKETS V E VEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EVVVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTE corresponding to amino acids 1 - 1527 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 1527 of HUMTEN PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PKPQLGTLIFSNITPKSFNMSWTTQAGLFAKIVINVSDAHSLHESQQFTVSGDAKQAHIT
GLVENTGYDVSVAGTTLAGDPTRPLTAFVITGTQSEVLTCLTQREKEISHLKGKFNKNTI
FTANVYSLIFN corresponding to amino acids 1528 - 1658 of HUMTEN PEA-1 P6, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence PKPQLGTLIFSNITPKSFNMSWTTQAGLFAKIVINVSDAHSLHESQQFTVSGDAKQAHIT
GLVENTGYDVSVAGTTLAGDPTRPLTAFVITGTQSEVLTCLTQREKEISHLKGKFNKNTI
FTANVYSLIFN in HUMTEN PEA I P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P7, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPV VFNH VYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
IO VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
I S FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATiNAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
EVWAEVGWDALKLN WTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTVVDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVT corresponding to amino acids 1 - 1617 of TENA HUMAN V1, which also corresponds to amino acids 1 - 1617 of I-IUMTEN PEA-1 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence GISNQVSHLFLFLVPFCVICLPDRHDFNIFVHIPYLIHKCSLLFHLLPTLPLVICT
corresponding to amino acids 1618 - 1673 of HUMTEN PEA-1 P7, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA_1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GISNQVSHLFLFLVPFCVICLPDRHDFNIFVHIPYLIHKCSLLFHLLPTLPLVICT in HUMTEN PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P8, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTV SIYGVIQGYRTPVLSAEASTGETPNLG
EVWAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
T corresponding to amino acids 1 - 1525 of TENA HUMAN V l, which also corresponds to amino acids 1 - 1525 of HUMTEN PEA-1 P8, and a second amino acid sequence being at least 90 % homologous to TEAEPEVDNLLV SDATPDGFRLSWTADEGVFDNFVLKIRDTKKQSEPLEITLLAPERTRD
LTGLREATEYEIELYGISKGRRSQTVSAIATTAMGSPKEVIFSDITENSATVSWRAPTAQV
ESFRITYVPITGGTPSMVTVDGTKTQTRLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTA
LDGPSGLVTANITDSEALARWQPAIATVDSYVISYTGEKVPEITRTVSGNTVEYALTDLE
PATEYTLRIFAEKGPQKSSTITAKFTTDLDSPRDLTATEVQSETALLTWRPPRASVTGYL
LVYESVDGTVKEVIVGPDTTSYSLADLSPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYP
FPKDCSQAMLNGDTTSGLYTIYLNGDKAQALEVFCDMTSDGGGWIVFLRRKNGRENF
YQNWKAYAAGFGDRREEFWLGLDNLNKITAQGQYELRVDLRDHGETAFAVYDKFSV
GDAKTRYKLKVEGYSGTAGDSMAYHNGRSFSTFDKDTDSAITNCALSYKGAFWYRNC
HRVNLMGRYGDNNHSQGVNWFHWKGHEHSIQFAEMKLRPSNFRNLEGRRKRA
corresponding to amino acids 1617 - 2201 of TENA_HUMAN V 1, which also corresponds to amino acids 1526 - 2110 of HUMTEN PEA_1 P8, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimerie polypeptide encoding for an edge portion of HUMTEN PEA-1 P8, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise TT, having a structure as follows: a sequence starting from any of amino acid numbers 1525-x to 1525; and ending at any of amino acid numbers 1526+ ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P10, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCI-ILRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EVWAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVL corresponding to amino acids 1 - 1252 of ilo TENA HUMAN V 1, which also corresponds to amino acids I - 1252 of HUMTEN PEA-1 P10, and a second amino acid sequence being at least 90 %
homologous to TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKV EAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTVVDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTAD
IATTAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQT
RLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIAT
VDSYVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTD
LDSPRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADL
SPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGD
KAQALEVFCDMTSDGGGWIVFLRRKNGRENFYQNWKAYAAGFGDRREEFWLGLDNL
NKITAQGQYELRVDLRDHGETAFAVYDKFSVGDAKTRYKLKVEGYSGTAGDSMAYH
NGRSFSTFDKDTDSAITNCALSYKGAFWYRNCHRVNLMGRYGDNNHSQGVNWFHWK
GHEHSIQFAEMKLRPSNFRNLEGRRKRA corresponding to amino acids 1344 - 2201 of TENA HUMAN V1, which also corresponds to amino acids 1253 - 2110 of HUMTEN PEA-1 P10, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMTEN PEA-1 P10, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise LT, having a structure as follows: a sequence starting from any of amino acid numbers 1252-x to 1252; and ending at any of amino acid numbers 1253+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P13, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSV DLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPV SARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EVWAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEW
corresponding to amino acids 1 - 1343 ofTENA HUMAN V1, which also corresponds to amino acids 1 - 1343 of HUMTEN PEA_1 P13, and a second amino acid sequence being at least 90 % homologous to TAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQTRLV
KLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIATVDS
YVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTDLDS
PRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADLSPS
THYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGDKAQ
ALEVFCDMTSDGGGWIVFLRRKNGRENFYQNWKAYAAGFGDRREEFWLGLDNLNKIT
AQGQYELRVDLRDI-IGETAFAVYDKFSVGDAKTRYKLKVEGYSGTAGDSMAYHNGRS
FSTFDKDTDSAITNCALSYKGAFWYRNCHRVNLMGRYGDNNHSQGVNWFHWKGHEH
SIQFAEMKLRPSNFRNLEGRRKRA corresponding to amino acids 1708 - 2201 of TENA HUMAN V1, which also corresponds to amino acids 1344 - 1837 of HUMTEN PEA 1 P13, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMTEN PEA-1 P
13, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise VT, having a structure as follows: a sequence starting from any of amino acid numbers 1343-x to 1343; and ending at any of amino acid numbers 1344+ ((rr2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_I P14, comprising a first amino acid sequence being at least 90 % homologous to LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTG LAPGQEYEISLHI V KNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EVVVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFI VYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTV SWMASENAFDSFLVTV VDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTAD
EGVFDNFVLKIRDTKKQSEPLEITLLAPERTRDLTGLREATEYEIELYGISKGRRSQTVSA
IATTAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQT
RLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIAT
VDSYVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTD
LDSPRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADL
SPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGD
KAQALEVFCDMTSDGGGWIV corresponding to amino acids 1 - 2025 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 2025 of HUMTEN PEA-1 P14, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence STTRDCRALRPRGRGRGQSRGGEEGDLLLMHSDTPMCEALQDSACHTEALRNSLLNKR
MGNTLATF corresponding to amino acids 2026 - 2091 of HUMTEN PEA_1 P14, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA_1 P 14, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence STTRDCRALRPRGRGRGQSRGGEEGDLLLMHSDTPMCEALQDSACHTEALRNSLLNKR
MGNTLATF in HUMTEN PEA 1 P14.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN_PEA_1 P15, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKAS corresponding to amino acids 1 - 1070 of TENA_HUMAN V 1, which also corresponds to amino acids 1 - 1070 of HUMTEN PEA-1 P15, and a second amino acid sequence being at least 90 % homologous to TEAEPEVDNLLVSDATPDGFRLSWTADEGVFDNFVLKIRDTKKQSEPLEITLLAPERTRD
LTGLREATEYEIELYGISKGRRSQTVSAIATTAMGSPKEVIFSDITENSATVSWRAPTAQV
ESFRITYVPITGGTPSMVTVDGTKTQTRLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTA
LDGPSGLVTANITDSEALARWQPAIATVDSYVISYTGEKVPEITRTVSGNTVEYALTDLE
PATEYTLRIFAEKGPQKSSTITAKFTTDLDSPRDLTATEVQSETALLTWRPPRASVTGYL
LVYESVDGTVKEVIVGPDTTSYSLADLSPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYP
FPKDCSQAMLNGDTTSGLYTIYLNGDKAQALEVFCDMTSDGGGWIVFLRRKNGRENF
YQNWKAYAAGFGDRREEFWLGLDNLNKITAQGQYELRVDLRDHGETAFAVYDKFSV
GDAKTRYKLKVEGYSGTAGDSMAYHNGRSFSTFDKDTDSAITNCALSYKGAFWYRNC
HRVNLMGRYGDNNHSQGVNWFHWKGHEHSIQFAEMKLRPSNFRNLEGRRKRA
corresponding to amino acids 1617 - 2201 of TENA HUMAN V 1, which also corresponds to amino acids 1071 - 1655 of HUMTEN PEA-1 P15, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMTEN PEA-1 P15, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise ST, having a structure as follows: a sequence starting from any of amino acid numbers 1070-x to 1070; and ending at any of amino acid numbers 1071+ ((m2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_I P16, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
FTGPDCAELSCPNDCI-IGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLV VTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPS.QIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRV SQTDNSITLEWRNGKAAIDSYRI KYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKAS corresponding to amino acids I - 1070 of TENA HUMAN VI, which also corresponds to amino acids I - 1070 of HUMTEN PEA-1 P16, and a second amino acid sequence being at least 90 % homologous to TAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQTRLV
KLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIATVDS
YVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTDLDS
PRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADLSPS
THYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGDKAQ
ALEVFCDMTSDGGGWIVFLRRKNGRENFYQNWKAYAAGFGDRREEFWLGLDNLNKIT
AQGQYELRVDLRDHGETAFAVYDKFSVGDAKTRYKLKVEGYSGTAGDSMAYHNGRS
FSTFDKDTDSAITNCALSYKGAFWYRNCHRVNLMGRYGDNNHSQGVNWFHWKGHEH
SIQFAEMKLRPSNFRNLEGRRKRA corresponding to amino acids 1708 - 2201 of TENA_HUMAN V 1, which also corresponds to amino acids 1071 - 1564 of HUMTEN PEA_1 P16, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMTEN PEA_1 P16, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise ST, having a structure as follows: a sequence starting from any of amino acid numbers 1070-x to 1070; and ending at any of amino acid numbers 107 I+ ((rr2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P 17, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDP LDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLR.AVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EV VVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTP LSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTVVDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTAD
EGVFDNFVLKIRDTKKQSEPLEITLLAPERTRDLTGLREATEYEIELYGISKGRRSQTVSA
IATTAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQT
RLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIAT
VDSYVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTD
LDSPRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADL
SPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGD
KAQALEVFCDMTSDGGGWIV corresponding to amino acids 1 - 2025 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 2025 of HUMTEN PEA-1 P17, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence TPWPTTMADPSPPLTRTQIQPSPTVLCPTKGLSGTGTVTVST corresponding to amino acids 2026 - 2067 of HUMTEN PEA-1 P17, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA-1 P17, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TPWPTTMADPSPPLTRTQIQPSPTVLCPTKGLSGTGTVTVST in HUMTEN PEA 1 P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P20, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCI-INRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTI DLTEDENQYSIGNLKPDTEYEV SLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVK.EDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQW VGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTV SIYGVIQGYRTPVLSAEASTGETPNLG
EVVVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTVVDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTAD
EGVFDNFVLKIRDTKKQSEPLEITLLAPERTRDLTGLREATEYEIELYGISKGRRSQTVSA
IATTAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQT
RLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIAT
VDSYVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTD
LDSPRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADL
SPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGD
KAQALEVFCDMTSDGGGWIVFLRRKNGRENFYQNWKAYAAGFGDRREEFWLG
corresponding to amino acids 1 - 2057 of TENA HUMAN V1, which also corresponds to amino acids 1 - 2057 of HUMTEN_PEA-1 P20, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NAALHVYI
corresponding to amino acids 2058 - 2065 of I-IUMTEN PEA-l P20, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN_PEA_1 P20, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NAALHVYI in HUMTEN PEA 1 P20.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P26, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPWFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EVVVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGL.K
AATHYTITIRGVTQDFSTTPLSVEVLTEEVP DMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTVVDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTAD
EGVFDNFVLKIRDTKKQSEPLEITLLAPERTRDLTGLREATEYEIELYGISKGRRSQTVSA
IATT corresponding to amino acids 1 - 1708 of TENA HUMAN V1, which also corresponds to amino acids 1 - 1708 of HUMTEN PEA-1 P26, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence 1 S GTVNKQERTEKSHDSGVFFSQG corresponding to amino acids 1709 - 1730 of HUMTEN PEA-1 P26, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA 1 P26, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GTVNKQERTEKSHDSGVFFSQG in HUMTEN PEA_1 P26.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_1 P27, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLV VYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRA VDIPGLKAATPYTV SIYGVIQGYRTPVLSAEASTGETPNLG
EVWAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
T corresponding to amino acids 1 - 1344 of TENA HUMAN V1, which also corresponds to amino acids 1 - 1344 of HUMTEN PEA-1 P27, and a second amino acid sequence being at least.70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GI
corresponding to amino acids 1345 - 1346 of HUMTEN PEA-1 P27, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P28, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLV VYTPTHEGGLEMQFRVPGDQTSTIIQELEPGV EYFIRVFAI
LENKKSIPV SARVATYLPAPEGLKFKSIKETSVEVEW DPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRV SQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EWVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLT corresponding to amino acids 1 - 1253 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 1253 of HUMTEN PEA-1 P28, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GILDEFTNSLPPLCLCSGGIKALSCFKLGSAPTTLGKYQ corresponding to amino acids 1254 - 1292 of HUMTEN PEA-I P28, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA-1 P28, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GILDEFTNSLPPLCLCSGGIKALSCFKLGSAPTTLGKYQ in HUMTEN PEA_1 P28.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_1 P29, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRI-IKRQSGVNATLPEENQPVVFNHVYNIK
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVC.DEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKAST corresponding to amino acids 1 - 1071 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 1071 of HUMTEN PEA-1 P29, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GESALSFLQTLG corresponding to amino acids 1072 - 1083 of HUMTEN PEA-1 P29, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA-1 P29, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GESALSFLQTLG in HUMTEN PEA 1 P29.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_1 P30, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTG corresponding to amino acids 1 - 954 of TENA HUMAN V l, which also corresponds to amino acids 1 - 954 of HUMTEN PEA-1 P30, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence ELCISASLSQPALEGP corresponding to amino acids 955 - 970 of HUMTEN PEA-1 P30, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA-1 P30, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence ELCISASLSQPALEGP in HUMTEN PEA 1 P30.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_1_P31, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCI-ILRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLWTEVTE
I S ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTR corresponding to amino acids 1 - 802 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 802 of HUMTEN PEA_1 P31, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence EYHL corresponding to amino acids 803 - 806 of HUMTEN PEA-1 P31, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA_1 P31, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% ho mologous to the sequence EYHL
in HUMTEN PEA 1 P31.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P32, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVAT corresponding to amino acids 1 - 710 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 710 of HUMTEN PEA-I P32, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence CE corresponding to amino acids 711 - 712 of HUMTEN PEA-1 P32, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMOSTRO PEA-1 PEA-1 P21, comprising a first amino acid sequence being at least 90 % homologous to MRIAVICFCLLGITCAIPVKQADSGSSEEKQLYNKYPDAVATWLNPDPSQKQNLLAPQ
corresponding to amino acids 1 - 58 of OSTP HUMAN, which also corresponds to amino acids 1 - 58 of HUMOSTRO PEA-1 PEA-1 P21, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VFLNFS
corresponding to amino acids 59 - 64 of HUMOSTRO PEA_1 PEA-1 P21, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMOSTRO PEA 1 PEA-1 P21, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence VFLNFS in HUMOSTRO PEA 1 PEA 1 P21.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMOSTRO PEA-1 PEA-1 P25, comprising a first amino acid sequence being at least 90 % homologous to MRIAVICFCLLGITCAIPVKQADSGSSEEKQ corresponding to amino acids 1 - 31 of OSTP HUMAN, which also con-esponds to amino acids 1 - 3 l of HUMOSTRO PEA-1 PEA-1 P25, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence H
corresponding to amino acids 32 - 32 of HUMOSTRO PEA_1 PEA-1 P25, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMOSTRO PEA-1 PEA_1 P30, comprising a first amino acid sequence being at least 90 % homologous to MRIAVICFCLLGITCAIPVKQADSGSSEEKQ corresponding to amino acids 1 - 31 of OSTP HUMAN, which also corresponds to amino acids 1 - 31 of HUMOSTRO PEA_1 PEA-1 P30, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence VSIFYVFI
corresponding to amino acids 32 - 39 of HUMOSTRO PEA-1 PEA-1 P30, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMOSTRO PEA_1 PEA 1 P30, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence VSIFYVFI in HUMOSTRO PEA 1 PEA 1 P30.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H61775 P16, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPR1DPDYVG corresponding to amino acids 1 t - 93 of Q9P2J2, which also corresponds to amino acids 1 - 83 of I-161775 P16, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV corresponding to amino acids 84 - 152 of H61775 P16, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of H61775 P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV in H61775 P16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H61775 P 16, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSWGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 1 - 83 of AAQ88495, which also corresponds to amino acids 1 - 83 of H61775 P16, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV corresponding to amino acids 84 - 152 of H61775 P16, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of H61775 P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV in H61775 P16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H61775 P17, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 1 I - 93 of Q9P2J2, which also corresponds to amino acids 1 - 83 of H61775_P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H61775 P17, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 1 - 83 of AAQ88495, which also corresponds to amino acids 1 - 83 of H61775 P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P2, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PHSGPAAAFIRRRGWWPGPRCA corresponding to amino acids 1 - 22 of HSAPHOL P2, second amino acid sequence being at least 90 % homologous to PATPRPLSWLRAPTRLCLDGPSPVLCA corresponding to amino acids 1 - 27 of AAH21289, which also corresponds to amino acids 23 - 49 of HSAPHOL P2, and a third amino acid sequence being at least 90 % homologous to EKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFLGDGMGVSTVTAARILKGQL
HHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLCGVKANEGTVGVSAAT
ERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAAYAHSADRDWYSDNE
MPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDVEYESDEKARGTRLD
GLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEPGDMQYELNRNNVT
DPSLSEM VWAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALHEAVEMDRAIGQAG
SLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYK
VVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQN
YVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 83 - 586 of AAH21289, which also corresponds to amino acids 50 -553 of I-ISAPHOL P2, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of HSAPHOL P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence PHSGPAAAFIRRRGWWPGPRCA of HSAPHOL P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of I-ISAPHOL P2, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise AE, having a structure as follows: a sequence starting from any of amino acid numbers 49-x to 50; and ending at any of amino acid numbers 50+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL_P2, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PHSGPAAAFIRRRGWWPGPRCAPATPRPLSWLRAPTRLCLDGPSPVLCA corresponding to amino acids 1 - 49 of HSAPHOL P2, second amino acid sequence being at least homologous to EKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFLGDGMGVSTVTAARILKGQL
HHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLCGVKANEGTVGVSAAT
ERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAAYAHSADRDWYSDNE
MPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDVEYESDEKARGTRLD
GLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEPGDMQYELNRNNVT
DPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALHEAVEMDRAIGQAG
SLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYK
WGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQN
YVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 21 - 524 of PPBT HUMAN, which also corresponds to amino acids 50 -553 of HSAPE-lOL-P2, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of HSAPHOL P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence PHSGPAAAFIRRRGWWPGPRCAPATPRPLSWLRAPTRLCLDGPSPVLCA of HSAPHOL P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P2, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise AE, having a structure as follows: a sequence starting from any of amino acid numbers 49-x to S0; and ending at any of amino acid numbers 50+ ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P3, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVP corresponding to amino acids 63 - 82 of AAH21289, which also corresponds to amino acids 1 - 20 of HSAPHOL P3, and a second amino acid sequence being at least 90 % homologous to GMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYL
CGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSA
AYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTD
VEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFE
PGDMQYELNRNNVTDPSLSEMV VVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQAL
HEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKK
PFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKG
PMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSV
LF corresponding to amino acids 123 - 586 of AAH21289, which also corresponds to amino acids 21 - 484 of HSAPHOL P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P3, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise PG, having a structure as follows: a sequence starting from any of amino acid numbers 20-x to 20; and ending at any of amino acid numbers 21+ ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P3, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVP corresponding to amino acids 1 - 20 of PPBT HUMAN, which also corresponds to amino acids 1 - 20 of HSAPHOL P3, and a second amino acid sequence being at least 90 % homologous to GMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYL
CGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSA
AYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTD
VEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFE
PGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQAL
HEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKK
PFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKG
PMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSV
LF corresponding to amino acids 61 - 524 of PPBT HUMAN, which also corresponds to amino acids 21 - 484 of HSAPHOL P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P3, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise PG, having a structure as follows: a sequence starting from any of amino acid numbers 20-x to 20; and ending at any of amino acid numbers 21+ ((n-2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P4, comprising a first amino acid sequence being at least 90 % homologous to MGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLC
GVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAA
YAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDV
EYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEP
GDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALH
EAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKP
FTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGP
MAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVL
F corresponding to amino acids 124 - 586 of AAH21289, which also corresponds to amino acids 1 - 463 of HSAPHOL P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P4, comprising a first amino acid sequence being at least 90 % homologous to MGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLC
GVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAA
YAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDV
EYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEP
GDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALH
EAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKP
FTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGP
MAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVL
F corresponding to amino acids 62 - 524 of PPBT HUMAN, which also corresponds to amino acids 1 - 463 of HSAPHOL P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P5, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
GLFEPGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKA
KQALHEAVEM corresponding to amino acids 63 - 417 of AAH21289, which also corresponds to amino acids 1 - 355 of HSAPHOL P5, and a second amino acid sequence being at least 90 homologous to DHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVD
YAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIG
ANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 440 - 586 of AAH21289, which also corresponds to amino acids 356 - 502 of HSAPHOL PS, wherein said 1 S first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P5, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise MD, having a structure as follows: a sequence starting from any ofamino acid numbers 355-x to 355; and ending at any of amino acid numbers 356+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHO L P5, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
GLFEPGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHI-IEGKA
KQALHEAVEM corresponding to amino acids 1 - 355 of .PPBT HUMAN, which also corresponds to amino acids 1 - 355 of HSAPHOL_P5, and a second amino acid sequence being at least 90 % homologous to DHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVD
YAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIG
ANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 377 - 524 of PPBT HUMAN, which also corresponds to amino acids 356 - 502 of HSAPHOL P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P5, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise MD, having a structure as follows: a sequence starting from any of amino acid numbers 355-x to 355; and ending at any of amino acid numbers 356+ ((rr2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P6, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
corresponding to amino acids 63 - 349 of AAH21289, which also corresponds to amino acids 1 287 of HSAPHOL P6, and a second amino acid sequence being at least 90 %
homologous to GGRIDHGHHEGKAKQALHEAVEMDRAIGQAGSLTSSEDTLTWTADHSHVFTFGGYTP
RGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAV
PLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAG
SLAAGPLLLALALYPLSVLF corresponding to amino acids 395 - 586 of AAH21289, which also corresponds to amino acids 288 - 479 of i-1SAPHOL P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P6, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise LG, having a structure as follows: a sequence starting from any of amino acid numbers 287-x to 287; and ending at any of amino acid numbers 288+ ((rr2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P6, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
corresponding to amino acids 1 - 287 of PPBT HUMAN, which also corresponds to amino acids 1 - 287 of HSAPHOL P6, and a second amino acid sequence being at least homologous to GGRIDHGHHEGKAKQALHEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTP
RGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAV
PLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAG
SLAAGPLLLALALYPLSVLF corresponding to amino acids 333 - 524 of PPBT HUMAN, which also corresponds to amino acids 288 - 479 of HSAPHOL Pti, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P6, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise LG, having a structure as follows: a sequence starting from any of amino acid numbers 287-x to 287; and ending at any of amino acid numbers 288+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P7, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYK corresponding to amino acids 63 -326 of AAH21289, which also corresponds to amino acids 1 - 264 of HSAPHOL P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, 1 S more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence LPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP
corresponding to amino acids 265 - 306 of HSAPHOL P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence LPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP in HSAPHOL P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P7, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPR corresponding to amino acids 1 - 262 of PPBT HUMAN, which also corresponds to amino acids 1 - 262 of HSAPHOL-P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence YKLPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP
corresponding to amino acids 263 - 306 of I-ISAPHOL P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence YKLPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP in HSAPHOL P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P7, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYK corresponding to amino acids 1 -264 of 075090, which also corresponds to amino acids 1 - 264 of HSAPHOL P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence LPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP corresponding to amino acids 265 - 306 of HSAPHOL P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence LPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP in HSAPHOL P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P8, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKI-ISHFIWNRTELLTLDPHNVDYLL
G corresponding to amino acids 63 - 350 of AAH21289, which also corresponds to amino acids 1 - 288 of HSAPHOL P8, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP corresponding to amino acids 289 - 316 of HSAPHOL P8, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P8, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP in HSAPHOL P8.
According to preferred embodiments of the present invention, there is provided an isolated chimerie polypeptide encoding for HSAPHOL P8, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
G corresponding to amino acids 1 - 288 of PPBT HUMAN, which also corresponds to amino acids 1 - 288 of HSAPHOL P8, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP corresponding to amino acids 289 - 316 of HSAPHOL P8, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P8, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP in HSAPHOL P8.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P8, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
G corresponding to amino acids 1 - 288 of 075090, which also corresponds to amino acids 1 -288 of HSAPHOL P8, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP corresponding to amino acids 289 - 316 of HSAPHOL P8, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P8, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP in HSAPHOL P8.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T10888 PEA_1 P2, comprising a first amino acid sequence being at least 90 % homologous to MGPPSAPPCRLHVPWKEVLLTASLLTFWNPPTTAKLTIESTPFNVAEGKEVLLLAHNLP
QNRIGYSWYKGERVDGNSLIVGYVIGTQQATPGPAYSGRETIYPNASLLIQNVTQNDTG
FYTLQVIKSDLVNEEATGQFHVYPELPKPSISSNNSNPVEDKDAVAFTCEPEVQNTTYL
WWVNGQSLPVSPRLQLSNGNMTLTLLSVKRNDAGSYECEIQNPASANRSDPVTLNVLY
YMCQAHNSATGLNRTTVTMITVS corresponding to amino acids 1 - 319 of CEA6 HUMAN, which also corresponds to amino acids 1 - 319 of T10888 PEA_1 P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DWTRP corresponding to amino acids 320 - 324 of T10888 PEA_1 P2, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T10888 PEA_I P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DWTRP in T10888 PEA 1 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T10888 PEA_I P4, comprising a first amino acid sequence being at least 90 % homologous to MGPPSAPPCRLHVPWKEVLLTASLLTFWNPPTTAKLTIESTPFNVAEGKEVLLLAHNLP
QNRIGYSWYKGERVDGNSLIVGYVIGTQQATPGPAYSGRETIYPNASLLIQNVTQNDTG
WWVNGQSLPVSPRLQLSNGNMTLTLLSVKRNDAGSYECEIQNPASANRSDPVTLNVL
corresponding to amino acids 1 - 234 of GEA6 HUMAN, which also corresponds to amino acids 1 - 234 of T10888 PEA I P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence LLLSSQLWPPSASRLECWPGWL corresponding to amino acids 235 - 256 of T10888 PEA_1 P4, wherein said first and second amino acid sequences are contiguois and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T10888 PEA-1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence LLLSSQLWPPSASRLECWPGWL in T10888 PEA 1 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T10888 PEA-1 P4, comprising a first amino acid sequence being at least 90 % homologous to MGPPSAPPCRLHVPWKEVLLTASLLTFWNPPTTAKLTIESTPFNVAEGKEVLLLAHNLP
QNRIGYSWYKGERVDGNSLIVGYVIGTQQATPGPAYSGRETIYPNASLLIQNVTQNDTG
FYTLQVIKSDLVNEEATGQFHVYPELPKPSISSNNSNPVEDKDAVAFTCEPEVQNTTYL
W WVNGQSLPVSPRLQLSNGNMTLTLLSVKRNDAGSYECEIQNPASANRSDPVTLNVL
corresponding to amino acids 1 - 234 of Q13774, which also corresponds to amino acids 1 - 234 of T10888 PEA-1 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence LLLSSQLWPPSASRLECWPGWL
corresponding to amino acids 235 - 256 of T10888 PEA_1 P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T10888 PEA-1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence LLLSSQLWPPSASRLECWPGWL in T10888 PEA 1 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T10888 PEA-1 P5, comprising a first amino acid sequence being at least 90 % homologous to MGPPSAPPCRLHVPWKEVLLTASLLTFWNPPTTAKLTIESTPFNVAEGKEVLLLAHNLP
QNRIGYSWYKGERVDGNSLIVGYVIGTQQATPGPAYSGRETIYPNASLLIQNVTQNDTG
FYTLQVIKSDLVNEEATGQFHVYPELPKPSISSNNSNPVEDKDAVAFTCEPEVQNTTYL
WWVNGQSLPVSPRLQLSNGNMTLTLLSVKRNDAGSYECEIQNPASANRSDPVTLNVLY
GPDVPTISPSKANYRPGENLNLSCHAASNPPAQYSWFINGTFQQSTQELFIPNITVNNSGS
YMCQAHNSATGLNRTTVTMITVSG corresponding to amino acids 1 - 320 of CEA6 HUMAN, which also corresponds to amino acids 1 - 320 of T 10888 PEA-1 P5, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide havingthe sequence VVFCFLISHV corresponding to amino acids 321 - 390 of T10888 PEA-1 P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T10888 PEA-1 P5, comprising a polypeptide being at. least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KWIHEALASHFQVESGSQRRARKKFSFPTCVQGAHANPKFSPEPSQFTSADSFPLVFLFF
WFCFLISHV in T10888 PEA 1 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T10888 PEA-1 P6, comprising a first amino acid sequence being at least 90 % homologous to MGPPSAPPCRLHVPWKEVLLTASLLTFWNPPTTAKLTIESTPFNVAEGKEVLLLAHNLP
QNRIGYSWYKGERVDGNSLIVGYVIGTQQATPGPAYSGRETIYPNASLLIQNVTQNDTG
FYTLQVIKSDLVNEEATGQFHW
corresponding to amino acids 1 - 141 of CEA6 HUMAN, which also corresponds to amino acids 1 - 141 of T10888 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence REYFHMTSGCWGSVLLPTYGIVRPGLCLWPSLHYILYQGLDI
corresponding to amino acids 142 - 183 of T10888 PEA 1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T10888 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence REYFHMTSGCWGSVLLPTYGIVRPGLCLWPSLHYILYQGLDI in T10888 PEA_I P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for I-ISECADH P9, comprising a first amino acid sequence being at least 90 % homologous to CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASV SGIQAELLTFPNSSPGLRRQKRDW VIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEG corresponding to amino acids 1 -274 of Q9UII7, which also corresponds to amino acids 1 - 274 of HSECADH P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG
corresponding to amino acids 275 - 322 of HSECADH P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSECADH_P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG in HSECADH P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH_P9, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEG corresponding to amino acids I -274 of Q9UII8, which also corresponds to amino acids 1 - 274 of HSECADH P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG
corresponding to amino acids 275 - 322 of HSECADH P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of I-ISECADH P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG in HSECADH P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P9, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEG corresponding to amino acids 1 -274 of CAD1 HUMAN, which also corresponds to amino acids 1 - 274 of HSECADH
P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homobgous to a polypeptide having the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG corresponding to amino acids 275 - 322 of HSECADH P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSECADH P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG in HSECADH P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P13, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFTINRNTGVISVVTTGLDRESFPTYTLVVQAADLQGEGL
STTATAVITVTDTNDNPPIFNPTT corresponding to amino acids 1 - 379 of Q9UII7, which also corresponds to amino acids 1 - 379 of HSECADH P13, and a second amino acid sequence VIL corresponding to amino acids 380 - 382 of HSECADH P13, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P 13, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFTINRNTGVISVVTTGLDRESFPTYTLVVQAADLQGEGL
STTATAVITVTDTNDNPPIFNPTT corresponding to amino acids 1 - 379 of Q9UII8, which also corresponds to amino acids I - 379 of HSECADH P13, and a second amino acid sequence VIL corresponding to amino acids 380 - 382 of HSECADH P13, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P13, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRI-ILERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
S GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFTINRNTGVISVVTTGLDRESFPTYTLVVQAADLQGEGL
STTATAV1TVTDTNDNPPIFNPTT corresponding to amino acids 1 - 379 of CAD 1-HUMAN, which also corresponds to amino acids 1 - 379 ofHSECADH_P13, and a second amino acid sequence V1L corresponding to amino acids 380 - 382 of HSECADH P13, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P 14, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAV SSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFTINRNTGVISVVTTGLDRE corresponding to amino acids 1 - 336 of Q9UII7, which also corresponds to amino acids 1 - 336 of HSECADH P
14, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRGQEDPEGVEDKCVLAQSRGQSK.ILLGQLSVNTVMV
corresponding to amino acids 337 - 373 of HSECADH P14, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSECADH P14, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRGQEDPEGVEDKCVLAQSRGQSKILLGQLSVNTVMV in HSECADH P14.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P 14, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFT1NRNTGVISVVTTGLDRE corresponding to amino acids 1'- 336 of Q9UII8, which also corresponds to amino acids 1 - 336 of HSECADH
P14, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 94% and most preferably at least 95% homologous to a polypeptide having the sequence VRGQEDPEGVEDKCVLAQSRGQSKILLGQLSVNTVMV
corresponding to amino acids 337 - 373 of HSECADH P14, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSECADH P 14, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRGQEDPEGVEDKCVLAQSRGQSKILLGQLSVNTVMV in HSECADH P14.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P14, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFTINRNTGVISWTTGLDRE corresponding to amino acids 1 - 336 of CADI HUMAN, which also corresponds to amino acids I - 336 of HSECADH P14, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRGQEDPEGVEDKCVLAQSRGQSKILLGQLSVNTVMV
corresponding to amino acids 337 - 373 of HSECADH P14, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSECADH P14, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRGQEDPEGVEDKCVLAQSRGQSKILLGQLSVNTVMV in HSECADH P14.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for I-ISECADH P15, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASV SGIQAELLTFPNSSPGLRRQKRDW VIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYT corresponding to amino acids 1 - 229 of Q9UII7, which also corresponds to amino acids 1 -229 of HSECADH P15, and a second amino acid sequence VSIS corresponding to amino acids 230 -233 of HSECADH P15, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P15, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYT corresponding to amino acids 1 - 229 of Q9UII8, which also corresponds to amino acids 1 -229 of HSECADH P15, and a second amino acid sequence VSIS corresponding to amino acids 230 -233 of HSECADH P15, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P15, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYT corresponding to amino acids 1 - 229 of CAD1 HUMAN, which also corresponds to amino acids 1 -229 of HSECADH P15, and a second amino acid sequence VSIS corresponding to amino acids 230 -233 of HSECADH P15, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P5, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYK corresponding to amino acids 12 - 55 of GILT HUMAN, which also corresponds to amino acids 1 - 44 of T59832 P5, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VGTATGRAGWREQAPCRGTRLLLSPQTSQGKTRAPRGRCPCRVPGKTLFSSRRCGHTP
SVPFRFRIPHLRGAAASTRLVPPKGSMSAYCVLLGQELGSPFVAQGTSSAAGQGPPACIL
AATLDAFIPARAGLACLWDLLGRCPRG corresponding to amino acids 45 - 189 of T59832 P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P5, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VGTATGRAGWREQAPCRGTRLLLSPQTSQGKTRAPRGRCPCRVPGKTLFSSRRCGHTP
SVPFRFRIPHLRGAAASTRLVPPKGSMSAYCVLLGQELGSPFVAQGTSSAAGQGPPACIL
AATLDAFIPARAGLACLWDLLGRCPRG in T59832 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P7, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQ ALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNG corresponding to amino acids 12 - 223 of GILT HUMAN, which also corresponds to amino acids 1 - 212 of T59832 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRIFLALSLTLIVPWSQGWTRQRDQR corresponding to amino acids 213 - 238 of T59832 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRIFLALSLTLIVPWSQGWTRQRDQR in T59832 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P7, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNG corresponding to amino acids 1 - 212 of BAC98466, which also corresponds to amino acids 1 - 212 of T59832 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRIFLALSLTLIVPWSQGWTRQRDQR corresponding to amino acids 213 - 238 of T59832 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRIFLALSLTLIVPWSQGWTRQRDQR in T59832 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P7, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV corresponding to amino acids 1 - 90 of T59832 P7, and a second amino acid sequence being at least 90 % homologous to MEILNVTLVPYGNAQEQNVSGRWEFKCQHGEEECKFNKVEACVLDELDMELAFLTIVC
MEEFEDMERSLPLCLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYV
PWVTVNGVRIFLALSLTLIVPWSQGWTRQRDQR corresponding to amino acids 1 - 148 of BAC85622, which also corresponds to amino acids 91 - 238 of T59832_P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of T59832 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV of T59832 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P7, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNG corresponding to amino acids 1 - 212 of Q8WU77, which also corresponds to amino acids 1 - 212 of.T59832 P7, and a second amino acid sequence being at least 70~%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRIFLALSLTLIVPWSQGWTRQRDQR corresponding to amino acids 213 - 238 of T59832 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRIFLALSLTLIVPWSQGWTRQRDQR in T59832 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P9, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHE corresponding to amino acids 12 - 214 of GILT HUMAN, which also corresponds to amino acids 1 - 203 of T59832 P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR corresponding to amino acids 204 - 244 of T59832 P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR in T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P9, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHE corresponding to amino acids 1 - 203 of BAC98466, which also corresponds to amino acids 1 - 203 of T59832 P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR corresponding to amino acids 204 - 244 of T59832 P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR in T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P9, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV corresponding to amino acids 1 - 90 of T59832 P9, second amino acid sequence being at least 90 % homologous to MEILNVTLVPYGNAQEQNVSGRWEFKCQHGEEECKFNKVEACVLDELDMELAFLTIVC
MEEFEDMERSLPLCLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHE
corresponding to amino acids 1 - 113 of BAC85622, which also corresponds to amino acids 91 -203 of T59832 P9, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR corresponding to amino acids 204 - 244 of T59832 P9, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV of T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR in T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P9, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHE corresponding to amino acids 1 - 203 of Q8WU77, which also corresponds to amino acids 1 - 203 of T59832 P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR corresponding to amino acids 204 - 244 of T59832 P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR in T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P12, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVE corresponding to amino acids 12 - 141 of GILT HUMAN, which also corresponds to amino acids 1 - 130 of T59832 P12, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 173 - 261 of GILT HUMAN, which also corresponds to amino acids 131 - 219 of T59832 P12, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P12, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EC, having a structure as follows: a sequence starting from any of amino acid numbers 130-x to 130; and ending at any of amino acid numbers 131+ ((n-2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P12, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV corresponding to amino acids 1 - 90 of T59832 P12, second amino acid sequence being at least 90 % homologous to MEILNVTLVPYGNAQEQNVSGRWEFKCQHGEEECKFNKVE corresponding to amino acids 1 - 40 of BAC85622, which also corresponds to amino acids 91 - 130 of T59832 P 12, third amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNG
corresponding to amino acids 72 - 122 of BAC85622, which also corresponds to amino acids 131 - 181 of T59832 P12, and a fourth amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence KPLEDQTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 182 - 219 of T59832 P12, wherein said first, second, third and fourth amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of T59832 P 12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV of T59832 P12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding far an edge portion of T59832 P12, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EC, having a structure as follows: a sequence starting from any of amino acid numbers 130-x to 130; and ending at any of amino acid numbers 131+ ((rr2) _ x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KPLEDQTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK in T59832 P12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P12, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVE corresponding to amino acids 1 - 130 of Q8WU77, which also corresponds to amino acids I - 130 of T59832 P12, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 162 - 250 of Q8WU77, which also corresponds to amino acids 131 - 219 of T59832 P12, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P 12, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EC, having a structure as follows: a sequence starting from any of amino acid numbers 130-x to 130; and ending at any of amino acid numbers 131+ ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832_P18, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYK corresponding to amino acids 12 - 55 of GILT HUMAN, which also corresponds to amino acids 1 - 44 of T59832 P18, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 173 - 261 of GILT HUMAN, which also corresponds to amino acids 45 - 133 of T59832 P18, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P18, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise KC, having a structure as follows: a sequence starting from any of amino acid numbers 44-x to 44; and ending at any of amino acid numbers 45+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P18, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYK corresponding to amino acids 1 - 44 of Q8 WU77, which also corresponds to amino acids 1 - 44 of T59832 P I 8, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 162 - 250 of Q8WU77, which also corresponds to amino acids 45 - 133 of T59832 P18, wherein said first and second amino acid sequences are contiguous and in a sequential order.
1 S According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P18, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise KC, having a structure as follows: a sequence starting from any of amino acid numbers 44-x to 44; and ending at any of amino acid numbers 45+ ((n-2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P18, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYK corresponding to amino acids 1 - 44 of Q8NEI4, which also corresponds to amino acids 1 - 44 of T59832 P18, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPW VTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 162 - 250 of Q8NE14, which also corresponds to amino acids 45 - 133 of T59832 P18, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P18, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about SO amino acids in length, wherein at least two amino acids comprise KC, having a structure as follows: a sequence starting from any of amino acid numbers 44-x to 44; and ending at any of amino acid numbers 45+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMGRPSE P4, comprising a first amino acid sequence being at least 90 % homologous to MRGSELPLVLLALVLCLAPRGRAVPLPAGGGTVLTKMYPRGNHWAVGHLMGKKSTG
ESSSVSERGSLKQQLREYIRWEEAARNLLGLIEAKENRNHQPPQPKALGNQQPSWDSED
SSNFKDVGSKGK corresponding to amino acids 1 - 127 of GRP-HUMAN, which also corresponds to amino acids 1 - 127 of HUMGRPSE P4, and a second amino acid sequence being at least 90 % homologous to GSQREGRNPQLNQQ corresponding to amino acids 148 of GRP-HUMAN, which also corresponds to amino acids 128 - 141 of HUMGRPSE
P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMGRPSE P4, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise KG, having a structure as follows: a sequence starting from any of amino acid numbers 127-x to 127; and ending at any of amino acid numbers 128 + ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMGRPSE P5, comprising a first amino acid sequence being at least 90 % homologous to MRGSELPLVLLALVLCLAPRGRAVPLPAGGGTVLTKMYPRGNHWAVGHLMGKKSTG
ESSSVSERGSLKQQLREYIRWEEAARNLLGLIEAKENRNHQPPQPKALGNQQPSWDSED
SSNFKDVGSKGK corresponding to amino acids 1 - I 27 of GRP_HUMAN, which also corresponds to amino acids I - 127 of HUMGRPSE P5, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DSLLQVLNVKEGTPS corresponding to amino acids 128 - 142 of HUMGRPSE P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMGRPSE P5, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DSLLQVLNVKEGTPS in HUMGRPSE P5.
According to preferred embodiments of the present invention, there is provided an isolated chimerie polypeptide encoding for 811723 PEA_1 P6, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAGSPCRGLAPGREEQRALHKAGAVGGGVR
corresponding to amino acids 1 - 110 of 811723 PEA-1 P6, and a second amino acid sequence being at least 90 % homo logous to MYAQALLWGVLQRQAAAQHLHEHPPKLLRGHRVQERVDDRAEVEKRLREGEEDHV
RPEVGPRPVVLGFGRSHDPPNLVGHPAYGQCHNNQPWADTSRRERQRKEKHSMRTQ
corresponding to amino acids 1 - 112 of Q8IXM0, which also corresponds to amino acids 111 -222 of 811723 PEA_1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of Rl 1723 PEA_1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAGSPCRGLAPGREEQRALHKAGAVGGGVR of 811723 PEA 1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA_1 P6, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 1 - 83 of Q96AC2, which also corresponds to amino acids 1 - 83 of 811723 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ corresponding to amino acids 84 - 222 of 811723 PEA 1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLWGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPWLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ in 811723 PEA-1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA_1 P6, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 1 - 83 of Q8N2G4, which also corresponds to amino acids 1 - 83 of 811723 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGI-IRVQERVDDRAEVEKRLREGEEDI-IVRPEVGPRPVVLGFGRSI-IDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKI-ISMRTQ corresponding to amino acids 84 - 222 of 811723 PEA 1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ in 811723 PEA-1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA_1 P6, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 24 - 106 of BAC85518, which also corresponds to amino acids 1 - 83 of 811723 PEA_1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ corresponding to amino acids 84 - 222 of 811723 PEA_1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ in 811723 PEA-1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for Rl 1723 PEA_1 P7, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAG corresponding to amino acids 1 - 64 of Q96AC2, which also corresponds to amino acids 1 - 64 of 811723 PEA-1 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of R I 1723 PEA-1 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA_1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in 811723 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA_1 P7, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAG corresponding to amino acids 1 - 64 of Q8N2G4, which also corresponds to amino acids 1 - 64 ofRl 1723 PEA-1 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of 811723 PEA-1 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in 811723 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P7, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWVLG corresponding to amino acids 1 - 5 of 811723 PEA-1 P7, second amino acid sequence being at least 90 % homologous to IAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEVMEQSAG
corresponding to amino acids 22 - 80 of BAC85273, which also corresponds to amino acids 6 -64 of RI 1723 PEA-1 P7, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of 811723 PEA-1 P7, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of 811723 PEA-1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MWVLG of 811723 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA_1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in 811723 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA_1 P7, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAG corresponding to amino acids 24 - 87 of BAC85518, which also corresponds to amino acids 1 - 64 of 811723-PEA-1 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of 811723 PEA-1 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of Rl 1723 PEA-1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in 811723 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P 13, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids I - 63 of Q96AC2, which also corresponds to amino acids 1 - 63 of 811723 PEA_1 P13, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DTKRTNTLLFEMRHFAKQLTT corresponding to amino acids 64 - 84 of 811723 PEA-1 P13, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P13, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DTKRTNTLLFEMRHFAKQLTT in 811723 PEA-1 P13.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P10, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids 1 - 63 of Q96AC2, which also corresponds to amino acids 1 - 63 of Rl 1723 PEA_1 P 10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of 811723 PEA_1 P10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in 811723 PEA 1 P10.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P10, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids 1 - 63 of Q8N2G4, which also corresponds to amino acids 1 - 63 of 811723 PEA-1 P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of 811723 PEA-1 P10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in 811723 PEA 1 P10.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P10, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWVLG corresponding to amino acids I - 5 of 811723 PEA-1 P10, second amino acid sequence being at least 90 % homologous to IAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVN VQDMCQKEVMEQSA
correspond ing to amino acids 22 - 79 of BAC85273, which also corresponds to amino acids 6 -63 of 811723 PEA-1 P 10, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of Rl 1723 PEA-1 P10, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of 811723 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MWVLG of 811723 PEA 1 P10.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in 811723 PEA-1 PIO.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P10, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids 24 - 86 of BAC85518, which also corresponds to amino acids 1 - 63 of 811723 PEA-1 P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of R 1 1723 PEA-I-P 10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in 811723 PEA-1 P10.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for D56406 PEA-1 P2, comprising a first amino acid sequence being at least 90 % homologous to MMAGMKIQLVCMLLLAFSSWSLCSDSEEEMKALEADFLTNMHTSKISKAHVPSWKMT
LLNVCSLVNNLNSPAEETGEVHEEELVARRKLPTALDGFSLEAMLTIYQLHKICHSRAF
QHWE corresponding to amino acids 1 - 120 of NEUT HUMAN, which also corresponds to amino acids 1 - 120 of D56406 PEA-1 P2, second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence ARWLTPVIPALWEAETGGSRGQEMETIPANT corresponding to amino acids 121 - 151 of D56406 PEA-1 P2, and a third amino acid sequence being at least 90 %
homologous to LIQEDILDTGNDKNGKEEVIKRKIPYILKRQLYENKPRRPYILKRDSYYY corresponding to amino acids 121 - 170 of NEUT HUMAN, which also corresponds to amino acids 152 - 201 of D56406 PEA_1 P2, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of D56406_PEA_1 P2, comprising an amino acid sequence being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence encoding for ARWLTPVIPALWEAETGGSRGQEMETIPANT, corresponding to D56406 PEA 1 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for D56406 PEA-1 P5, comprising a first amino acid sequence being at least 90 % homologous to MMAGMKIQLVCMLLLAFSSWSLC
corresponding to amino acids 1 - 23 of NEUT_1-IUMAN, which also corresponds to amino acids 1 - 23 of D56406 PEA-1-P5, and a second amino acid sequence being at least 90 homologous to SEEEMKALEADFLTNMI-ITSKISKAHVPSWKMTLLNVCSLVNNLNSPAEETGEVHEEEL
VARRKLPTALDGFSLEAMLTIYQLHKICHSRAFQHWELIQEDILDTGNDKNGKEEVIKR
KIPYILKRQLYENKPRRPYILKRDSYYY corresponding to amino acid s 26 - 170 of NEUT HUMAN, which also corresponds to amino acids 24 - 168 of D56406 PEA-1 P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of D56406 PEA_1 P5, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise CS, having a I S structure as follows: a sequence starting from any of amino acid numbers 23-x to 24; and ending at any of amino acid numbers + ((rr2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for D56406 PEA-I P6, comprising a first amino acid sequence being at least 90 % homologous to MMAGMKIQLVCMLLLAFSSWSLCSDSEEEMKALEADFLTNMHTSK corresponding to amino acids 1 - 45 of NEUT HUMAN, which also corresponds to amino acids 1 - 45 of D56406 PEA-1 P6, and a second amino acid sequence being at least 90 %
homologous to LIQEDILDTGNDKNGKEEVIKRKIPYILKRQLYENKPRRPYILKRDSYYY corresponding to amino acids 121 - 170 of NEUT_HUMAN, which also corresponds to amino acids 46 -95 of D56406 PEA 1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of D56406 PEA_1 P6, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise KL, having a structure as follows: a sequence starting from any of amino acid numbers 45-x to 46; and ending at any of amino acid numbers 46+ ((n-2) - x), in which x varies from 0 to r~2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H53393 PEA-1 P2, comprising a first amino acid sequence being at least 90 % homologous to MRTYRYFLLLFWVGQPYPTLSTPLSKRTSGFPAKKRALELSGNSKNELNRSKRSWMWN
QFFLLEEYTGSDYQYVGKLHSDQDRGDGSLKYILSGDGAGDLFIINENTGDIQATKRLD
QVTATDADDPTYGNSAKVVYSILQGQPYFSVESETGIIKTALLNMDRENREQYQWIQA
KDMGGQMGGLSGTTTVNITLTDVNDNPPRFPQSTYQFKTPESSPPGTPIGRIKASDADV
FLYLGPFKDSATVRIVVEDVDEPPVFSKLAYILQIREDAQINTTIGSVTAQDPDAARNPV
KYSVDRHTDMDRIFNIDSGNGSIFTSKLLDRETLLWHNITVIATEINNPKQSSRVPLYIKV
LDVNDNAPEFAEFYETFVCEKAKADQLIQTLHAVDKDDPYSGHQFSFSLAPEAASGSNF
TIQDNK corresponding to amino acids 1 - 543 of CAD6_HUMAN, which also corresponds to amino acids 1 - 543 of H53393 PEA-I P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GK
corresponding to amino acids 544 - 545 of H53393 PEA 1 P2, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H53393 PEA_1 P3, comprising a first amino acid sequence being at least 90 % homologous to MRTYRYFLLLFWGQPYPTLSTPLSKRTSGFPAKKRALELSGNSKNELNRSKRSWMWN
REEKPWILRAQAINRRTGRPVEPESEFIIKIHDINDNEPIFTKEVYTATVPEMSDVGTFW
QVTATDADDPTYGNSAKVVYSILQGQPYFSVESETGIIKTALLNMDRENREQYQWIQA
KDMGGQMGGLSGTTTVNITLTDVNDNPPRFPQSTYQFKTPESSPPGTPIGRIKASDADV
GENAEIEYSITDGEGLDMFDVITDQETQEGIITVKKLLDFEKKKVYTLKVEASNPWEPR
FLYLGPFKDSATVRIWEDVDEPPVFSKLAYILQIREDAQINTTIGSVTAQDPDAARNPV
LDVNDNAPEFAEFYETFVCEKAKADQ corresponding to amino acids 1 - 504 of CAD6 HUMAN, which also corresponds to amino acids 1 - 504 of H53393 PEA-1 P3, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence RFGFSLS corresponding to amino acids 505 - S 11 of H53393 PEA_I P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of H53393 PEA-1 P3, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence RFGFSLS
in H53393 PEA 1 P3.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H53393 PEA-1 P6, comprising a first amino acid sequence being at least 90 % homologous to MRTYRYFLLLFWVGQPYPTLSTPLSKRTSGFPAKKRALELSGNSKNELNRSKRSWMWN
QFFLLEEYTGSDYQYVGKLHSDQDRGDGSLKYILSGDGAGDLFIINENTGDIQATKRLD
REEKPVYILRAQAINRRTGRPVEPESEFIIKIHDINDNEPIFTKEVYTATVPEMSDVGTFVV
QVTATDADDPTYGNSAKVVYSILQGQPYFSVESETGIIKTALLNMDRENREQYQWIQA
KDMGGQMGGLSGTTTVNITLTDVNDNPPRFPQSTYQFKTPESSPPGTPIGRIKASDADV
GENAEIEYSITDGEGLDMFDVITDQETQEGIITVKK corresponding to amino acids 1 - 333 of CAD6 HUMAN, which also corresponds to amino acids I - 333 of H53393 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VMPLLKHHTE corresponding to amino acids 334 -343 of H53393 PEA-1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of H53393 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VMPLLKHHTE in H53393 PEA 1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSU40434 PEA-1 P12, comprising a first amino acid sequence being at least 90 % homologous to MALPTARPLLGSCGTPALGSLLFLLFSLGWVQPSRTLAGETGQEAAPLDGVLANPPNISS
LSPRQLLGFPCAEVSGLSTERVRELAVALAQKNVKLSTEQLRCLAHRLSEPPEDLDALP
LDLLLFLNPDAFSGPQACTRFFSRITKANV DLLPRGAPERQRLLPAALACWGVRGSLLS
EADVRALGGLACDLPGRFVAESAEVLLPRLVSCPGPLDQDQQEAARAALQGGGPPYGP
PSTWSVSTMDALRGLLPVLGQPIIRSIPQGIVAAWRQRSSRDPSWRQPERTILRPRFRRE
VEKTACPSGKKAREIDESLIFYKKWELEACVDAALLATQMDRVNAIPFTYEQLDVLKH
KLDELYPQGYPESVIQHLGYLFLKMSPEDIRKWNVTSLETLKALLEVNKGHEMSPQVA
TLIDRFVKGRGQLDKDTLDTLTAFYPGYLCSLSPEELSSVPPSSIW corresponding to amino acids 1 - 458 of Q14859, which also corresponds to amino acids 1 - 458 of HSU40434 PEA 1 P 12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSU40434 PEA-1 P12, comprising a first amino acid sequence being at least 90 % homologous to MALPTARPLLGSCGTPALGSLLFLLFSLGWVQPSRTLAGETGQ corresponding to amino acids I - 43 of Q9BTR2, which also corresponds to amino acids I - 43 of HSU40434 PEA-1 P 12, second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence E corresponding to amino acids 44 - 44 of HSU40434 PEA-1 P12, and a third amino acid sequence being at least 90 %
homologous to AAPLDGVLANPPNISSLSPRQLLGFPCAEVSGLSTERVRELAVALAQKNVKLSTEQLRC
LAHRLSEPPEDLDALPLDLLLFLNPDAFSGPQACTRFFSRITKANVDLLPRGAPERQRLL
PAALACWGVRGSLLSEADVRALGGLACDLPGRFVAESAEVLLPRLVSCPGPLDQDQQE
AARAALQGGGPPYGPPSTWSVSTMDALRGLLPVLGQPIIRSIPQGIVAAWRQRSSRDPS
WRQPERTILRPRFRREVEKTACPSGKKAREIDESLIFYKKWELEACVDAALLATQMDRV
NAIPFTYEQLDVLKHKLDELYPQGYPESVIQHLGYLFLKMSPEDIRKWNVTSLETLKAL
LEVNKGHEMSPQVATLIDRFVKGRGQLDKDTLDTLTAFYPGYLCSLSPEELSSVPPSSIW
corresponding to amino acids 44 - 457 of Q9BTR2, which also corresponds to amino acids 45 -458 ofHSU40434 PEA-1 P12, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of HSU40434 PEA_ 1 P 12, comprising an amino acid sequence being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at.least about 90% and most preferably at least about 95%
homologous to the sequence encoding for E, corresponding to HSU40434 PEA 1 P12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P2, comprising a first amino acid sequence being at least 90 % homologous to MLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNRTFIWD
VKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATVVRIGTFCSNGTVSRIKMQ
EGVKMALHLPWFHPRNVSGFSIANRSSIKRLCIIESVFEGEGSATLMSANYPEGFPEDEL
MTWQFVVPAHLRASVSFLNFNLSNCERKEERVEYYIPGSTTNPEVFKLEDKQPGNMAG
NFNLSLQGCDQDAQSPGILRLQFQVLVQHPQNES corresponding to amino acids 67 - 341 of Q8WU91, which also corresponds to amino acids 1 - 275 of M77904 P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NKIYVVDLSNERAMSLTIEPRPVKQSRKFVPGCFVC LESRTCSSNLTLTSGSKHKISFLCD
DLTRLWMNVEKTISCTDHRYCQRKSYSLQVPSDILHLPVELHDFSWKLLVPKDRLSLVL
VPAQKLQQHTHEKPCNTSFSYLVASAIPSQDLYFGSFCPGGSIKQIQVKQNISVTLRTFAP
SFQQEASRQGLTVSFIPYFKEEGVFTVTPDTKSKVYLRTPNWDRGLPSLTSVSWNISVPR
DQVACLTFFKERSGWCQTGRAFMIIQEQRTRAEEIFSLDEDVLPKPSFHHHSFWVNISN
CSPTSGKQLDLLFSVTLTPRTVDLTVILIAAVGGGVLLLSALGLIICCVKKKKKKTNKGP
AVGIYNGNINTEMPRQPKKFQKGRKDNDSHVYAVIEDTMWGHLLQDSSGSFLQPEVD
TYRPFQGTMGVCPPSPPTICSRAPTAKLATEEPPPRSPPESESEPYTFSHPNNGDVSSKDT
DIPLLNTQEPMEPAE corresponding to amino acids 276 - 770 of M77904 P2, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NKIYVVDLSNERAMSLTIEPRPVKQSRKFVPGCFVCLESRTCSSNLTLTSGSKHKISFLCD
VPAQKLQQHTHEKPCNTSFSYLVASAIPSQDLYFGSFCPGGSIKQIQVKQNISVTLRTFAP
SFQQEASRQGLTVSFIPYFKEEGVFTVTPDTKSKVYLRTPNWDRGLPSLTSVSWNISVPR
DQVACLTFFKERSGVVCQTGRAFMIIQEQRTRAEEIFSLDEDVLPKPSFHHHSFWVNISN
CSPTSGKQLDLLFSVTLTPRTVDLTVILIAAVGGGVLLLSALGLIICCVKKKKKKTNKGP
AVGIYNGNINTEMPRQPKKFQKGRKDNDSHVYAVIEDTMVYGHLLQDSSGSFLQPEVD
TYRPFQGTMGVCPPSPPTICSRAPTAKLATEEPPPRSPPESESEPYTFSHPNNGDVSSKDT
DIPLLNTQEPMEPAE in M77904 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P2, comprising a first amino acid sequence being at least 90 % homologous to MLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNRTFIWD
VKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATV VRIGTFCSNGTVSRIKMQ
EGVKMALHLPWFHPRNVSGFSIANRSSIKRLCIIESVFEGEGSATLMSANYPEGFPEDEL
MTWQFVVPAHLRASVSFLNFNLSNCERKEERVEYYIPGSTTNPEVFKLEDKQPGNMAG
NFNLSLQGCDQDAQSPGILRLQFQVLVQHPQNESNKIYVVDLSNERAMSLTIEPRPVKQ
SRKFVPGCFVCLESRTCSSNLTLTSGSKHKISFLCDDLTRLWMNVEKTISCTDHRYCQR
KSYSLQVPSDILHLPVELHDFSWKLLVPKDRLSLVLVPAQKLQQHTHEKPCNTSFSYLV
ASAIPSQDLYFGSFCPGGSIKQIQVKQNISVTLRTFAPSFQQEASRQGLTVSFIPYFKEEGV
FTVTPDTKSKVYLRTPNWDRGLPSLTSVSWNISVPRDQVACLTFFKERSGVVCQTGRAF
MIIQEQRTRAEEIFSLDEDVLPKPSFHHHSFWVNISNCSPTSGKQLDLLFSVTLTPRTVDL
TVILIAAVGGGVLLLSALGLIICCVKKKKKKTNKGPAVGIYNGNINTEMPRQPKKFQKG
RKDNDSHVYAVIEDTMVYGHLLQDSSGSFLQPEVDTYRPFQGTMGVCPPSPPTICSRAP
TAKLATEEPPPRSPPESESEPYTFSHPNNGDVSSKDTDIPLLNTQEPMEPAE corresponding to amino acids 67 - 836 of Q96QU7, which also corresponds to amino acids 1 -770 of M77904 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P4, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGVSIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
KRHITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATV VRIGTFCSNGTVSR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKRLCIIESVFEGEGSATLMSANYPEGFP
EDELMTWQFVVPAHLRASVSFLNFNLSNCERKEERVEYYIPGSTTNPEVFKLEDKQPGN
MAGNFNLSLQGCDQDAQSPGILRLQFQVLVQHPQNES corresponding to amino acids 1 -341 of Q8WU91, which also corresponds to amino acids 1 - 341 of M77904 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NKIYWDLSNERAMSLTIEPRPVKQSRKFVPGCFVCLESRTCSSNLTLTSGSKHKISFLCD
DLTRLWMNVEKTISTPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLT
PVIPALWEAKAGGSLEVRSSRPAWPTW corresponding to amino acids 342 - 487 of M77904 P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail ofM77904 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NKIYWDLSNERAMSLTIEPRPVKQSRKFVPGCFVC LESRTCSSNLTLTSGSKHKISFLCD
DLTRLWMNVEKTISTPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLT
PVIPALWEAKAGGSLEVRSSRPAWPTW in M77904 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P4, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGVSIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
KRHITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATWRIGTFCSNGTVSR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKRLCIIESVFEGEGSATLMSANYPEGFP
EDELMTWQFVVPAHLRASVSFLNFNLSNCERKEERVEYYIPGSTTNPEVFKLEDKQPGN
MAGNFNLSLQGCDQDAQSPGILRLQFQVLVQHPQNESNKIYVVDLSNERAMSLTIEPRP
VKQSRKFVPGCFVCLESRTCSSNLTLTSGSKHKISFLCDDLTRLWMNVEKTIS
corresponding to amino acids I - 416 of Q9HSV8, which also corresponds to amino acids 1 -416 of M77904 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence TPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLTPVIPALWEAKAGGS
LEVRSSRPAWPTW corresponding to amino acids 417 - 487 of M77904 P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about I S 90% and most preferably at least about 95% homologous to the sequence TPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLTPVIPALWEAKAGGS
LEVRSSRPAWPTW in M77904 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P4, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGVSIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
KRI-IITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATVVRIGTFCSNGTVSR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKRLCIIESVFEGEGSATLMSANYPEGFP
EDELMTWQFWPAHLRASVSFLNFNLSNCERKEERVEYYIPGSTTNPEVFKLEDKQPGN
MAGNFNLSLQGCDQDAQSPGILRLQFQVLVQHPQNESNKIYVVDLSNERAMSLTIEPRP
VKQSRKFVPGCFVCLESRTCSSNLTLTSGSKHKISFLCDDLTRLWMNVEKTIS
corresponding to amino acids 1 - 416 of Q96QU7, which also corresponds to amino acids 1 -416 of M77904 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence TPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLTPVIPALWEAKAGGS
LEVRSSRPAWPTW corresponding to amino acids 417 - 487 of M77904 P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLTPVIPALWEAKAGGS
LEVRSSRPAWPTW in M77904 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P5, comprising a first amino acid sequence being at least 90 % homologous to MIIQEQRTRAEEIFSLDEDVLPKPSFHHHSFWVNISNCSPTSGKQLDLLFSVTLTPRTVDL
TVILIAAVGGGVLLLSALGLIICCVKKKKKKTNKGPAVGIYNGNINTEMPRQPKKFQKG
RKDNDSHVYAVIEDTMVYGHLLQDSSGSFL,QPEVDTYRPFQGTMGVCPPSPPTICSRAP
TAKLATEEPPPRSPPESESEPYTFSHPNNGDVSSKDTDIPLLNTQEPMEPAE corresponding to amino acids 606 - 836 of Q96QU7, which also corresponds to amino acids 1 -231 of M77904 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P5, comprising a first amino acid sequence being at least 90 % homologous to MIIQEQRTRAEEIFSLDEDVLPKPSFHHHSFWVNISNCSPTSGKQLDLLFSVTLTPRTVDL
TVILIAAVGGGVLLLSALGLIICCVKKKKKKTNKGPAVGIYNGNINTEMPRQPKKFQKG
RKDNDSHVYAVIEDTMVYGHLLQDSSGSFLQPEVDTYRPFQGTMGVCPPSPPTICSRAP
TAKLATEEPPPRSPPESESEPYTFSHPNNGDVSSKDTDIPLLNTQEPMEPAE corresponding to amino acids 419 - 649 of Q9H8C2, which also corresponds to amino acids 1 -231 of M77904 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P7, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGVSIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
18o KRHITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATVVRIGTFCSNGTVSR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKR corresponding to amino acids 1 - 219 of Q8WU91, which also corresponds to amino acids 1 - 219 ofM77904 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence EKAPPCYLIRLKHTRSSLF corresponding to amino acids 220 - 238 of M77904 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence EKAPPCYLIRLKHTRSSLF in M77904 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P7, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGVSIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
KRHITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATVVRIGTFCSNGTVSR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKR corresponding to amino acids 1 - 219 of Q9HSV8, which also corresponds to amino acids 1 - 219 of M77904 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence EKAPPCYLIRLKHTRSSLF corresponding to amino acids 220 - 238 of M77904 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence EKAPPCYLIRLKHTRSSLF in M77904 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P7, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGV SIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
KRHITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATV VRIGTFCSNGTV SR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKR corresponding to amino acids 1 - 219 of Q96QU7, which also corresponds to amino acids 1 - 219 of M77904 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence EKAPPCYLIRLKHTRSSLF corresponding to amino acids 220 - 238 of M77904 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence EKAPPCYLIRLKHTRSSLF in M77904 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 225299 PEA 2 P2, comprising a first amino acid sequence being at least 90 % homologous to MKSSGLFPFLVLLALGTLAPWAVEGSGKSFKAGVCPPKKSAQCLRYKKPECQSDWQCP
GKKRCCPDTCGIKCLDPVDTPNPTRRKPGKCPVTYGQCLMLNPPNFCEMDGQCKRDLK
CCMGMCGKSCVSPVK corresponding to amino acids 1 - 131 of ALK1 HUMAN, which also corresponds to amino acids 1 - 131 of 225299 PEA 2 P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GKQGMRAH corresponding to amino acids 132 - 139 of 225299 PEA 2 P2, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 225299 PEA 2 P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homobgous to the sequence GKQGMRAH in 225299 PEA 2 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 225299 PEA 2 P3, comprising a first amino acid sequence being at least 90 % homologous to MKSSGLFPFLVLLALGTLAPWAVEGSGKSFKAGVCPPKKSAQCLRYKKPECQSDWQCP
GKKRCCPDTCGIKCLDPVDTPNPTRRKPGKCPVTYGQCLMLNPPNFCEMDGQCKRDLK
CCMGMCGKSCVSPVK corresponding to amino acids 1 - 131 of ALKI HUMAN, which also corresponds to amino acids 1 - 131 of 225299 PEA 2 P3, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GEKRHHKQLRDQEVDPLEMRRHSAG corresponding to amino acids 132 - 156 of 225299 PEA 2 P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 225299 PEA 2 P3, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GEKRHHKQLRDQEVDPLEMRRHSAG in 225299 PEA 2 P3.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 225299 PEA 2 P7, comprising a first amino acid sequence being at least 90 % homologous to MKSSGLFPFLVLLALGTLAPWAVEGSGKSFKAGVCPPKKSAQCLRYKKPECQSDWQCP
GKKRCCPDTCGIKCLDPVDTPNP corresponding to amino acids 1 - 81 of ALKI HUMAN, which also corresponds to amino acids I - 81 of 225299 PEA 2 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence RGSLGSAQ corresponding to amino acids 82 - 89 of 225299 PEA 2-P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 225299 PEA 2 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence RGSLGSAQ in 225299 PEA 2 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 225299 PEA 2 PIO, comprising a first amino acid sequence being at least 90 % homologous to MKSSGLFPFLVLLALGTLAPWAVEGSGKSFKAGVCPPKKSAQCLRYKKPECQSDWQCP
GKKRCCPDTCGIKCLDPVDTPNPT corresponding to amino acids 1 - 82 ofALKI HUMAN, which also corresponds to amino acids 1 - 82 of 225299 PEA 2 P10.
According to preferred embodiments of the present invention, there is provided an antibody capable of specifically binding to an epitope of an amino acid sequence as described herein.
Optionally the amino acid sequence corresponds to a bridge, edge portion, tail, head or insertion as described herein.
Optionally the antibody is capable of differentiating between a splice variant having said epitope and a corresponding known protein.
According to preferred embodiments of the present invention, there is provided a kit for detecting ovarian cancer, comprising a kit detecting overexpression of a splice variant as described herein.
Optionally the kit comprises a NAT-based technology.
Optionally the kit further comprises at least one primer pair capable of selectively hybridizing to a nucleic acid sequence as described herein.
Optionally the kit further comprises at least one oligonucleotide capable of selectively hybridizing to a nucleic acid sequence as described herein.
Optionally the kit comprises an antibody as described herein.
Optionally the kit further comprises at least one reagent for performing an ELISA or a Western blot.
According to preferred embodiments of the present invention, there is provided a method for detecting ovarian cancer, comprising detecting overexpression of a splice variant as described herein.
Optionally detecting overexpression is performed with a NAT-based technology.
Optionally detecting overexpression is performed with an immunoassay.
Optionally the irmnunoassay comprises an antibody as described herein.
According to preferred embodiments of the present invention, there is provided a biomaiker capable of detecting ovarian cancer, comprising any of the above nucleic acid sequences or a fragment thereof, or any of the above amino acid sequences or a fragment thereof.
According to preferred embodiments of the present invention, there is provided a method for screening for ovarian cancer, comprising detecting ovarian cane er cells with a biomarker or an antibody or a method or assay as described herein.
According to preferred embodiments of the present invention, there is provided a method for diagnosing ovarian cancer, comprising detecting ovarian cancer cells with a biomarker or an antibody or a method or assay as described herein.
According to preferred embodiments of the present invention, there is provided a method for monitoring disease progression and/or treatment efficacy and/or relapse of ovarian cancer, comprising detecting ovarian cancer cells with a biomarker or an antibody or a method or assay as described herein.
According to preferred embodiments of the present invention, there is provided a method of selecting a therapy for ovarian cancer, comprising detecting ovarian cancer cells with a biomarker or an antibody or a method or assay as described herein and selecting a therapy according to said detection.
According to preferred embodiments of the present invention, preferably any of the above nucleic acid and/or amino acid sequences further comprises any sequence having at least about 70%, preferably at least about 80%, more preferably at least about 90%, most preferably at least about 95% homology thereto.
Unless otherwise noted, all experimental data relates to variants of the present invention, named according to the segment being tested (as expression was tested through RT-PCR as described).
All nucleic acid sequences and/or amino acid sequences shown herein as embodiments of the present invention relate to their isolated form, as isolated polynucleotides (including for all transcripts), oligonucleotides (including for all segments, amplicons and primers), peptides (including for all tails, bridges, insertions or heads, optionally including other antibody epitopes as described herein) and/or polypeptides (including for all proteins). It should be noted that oligonucleotide and polynucleotide, or peptide and polypeptide, may optionally be used interchangeably.
BRIEF DESCRIPTION OF DRAWINGS
Figure 1 is schematic summary of cancer biomarkers selection engine and the wet validation stages.
Figure 2. Schematic illustration, depicting grouping of transcripts of a given cluster based on presence or absence of unique sequence regions.
Figure 3 is schematic summary of quantitative real-time PCR analysis.
Figure 4 is schematic presentation of the oligonucleotide based microarray fabrication.
Figure 5 is schematic summary of the oligonucleotide based microarray experimental flow.
Figure 6 shows cancer and cell-line vs. normal tissue expression for .
Figure 7 shows expression of segment8 in H61775 in cancerous vs. norrcancerous tissues.
Figure 8 shows expression of segment8 in H61775 in normal tissues.
Figure 9 shows cancer and cell-line vs. normal tissue expression.
Figure 10 is a histogram showing over expression of T juncl 1-17 transcripts in cancerous ovary samples relative to the normal samples.
Figure 11 is a histogram showing expression of T juncl 1-17 transcripts in normal tissues.
Figure 12 shows cancer and cell-line vs. normal tissue expression.
Figure 13 is a histogram showing over expression of HUMGRPSEjunc3-7 transcripts in cancerous ovary samples relative to the normal samples.
Figure 14 is a histogram showing expression of HUMGRPSEjunc3-7 transcripts in normal tissues.
Figure 15 shows cancer and cell-line vs. normal tissue expression.
Figure 16 is a histogram showing over expression of 811723 segl3 transcripts in S cancerous ovary samples relative to the normal PM samples.
Figure 17 is a histogram showing expression of RI 1723 segl3 transcripts in normal tissue samples.
Figure 18 is a histogram showing over expression of R 11723 junc 11-18 transcripts in cancerous ovary samples relative to the normal samples.
Figure 19 is a histogram showing expression of 811723 juncl l-18 transcripts in normal tissue samples.
Figure 20 shows cancer and cell-line vs. normal tissue expression.
Figure 21 is a histogram showing over expression of H53393 segl3 transcripts in cancerous ovary samples relative to the normal samples.
I S Figure 22 is a histogram showing over expression of H53393 junc2l-22 transcripts in cancerous ovary samples relative to the normal samples.
Figure 23 shows cancer and cell-line vs. normal tissue expression.
Figure 24 shows cancer and cell-line vs. normal tissue expression.
Figure 25 shows cancer and cell-line vs. normal tissue expression.
Figure 26 is a histogram showing over expression of 225299 juncl3-14-21 transcripts in cancerous ovary samples relative to the normal samples.
Figures 27A and 27B are histograms showing over expression of 225299 seg20 transcripts in cancerous ovary samples relative to the normal samples (27A) or in normal tissues (27B).
Figures 28A and 28B are histograms showing over expression of 225299 seg23 transcripts in cancerous ovary samples relative to the normal samples (28A) or in normal tissues (28B).
Figure 29 shows cancer and cell-line vs. normal tissue expression.
Figure 30 is a histogram showing down regulation of T39971 june23-338 transcripts in cancerous ovary samples relative to the normal samples.
Figure 31 is a histogram showing expression of T39971 junc23-33R transcripts in normal tissues.
Figure 32 shows cancer and cell-line vs. normal tissue expression.
Figures 33A and 33B are histograms showing down regulation of 244808 junc8-11 transcripts in cancerous ovary samples relative to the normal samples (33A) or expression in normal tissues (33B).
Figure 34 shows cancer and cell-line vs. normal tissue expression.
Figure 35 shows cancer and cell-line vs. normal tissue expression.
Figure 36 shows cancer and cell-line vs. normal tissue expression.
Figure 37 shows cancer and cell-line vs. normal tissue expression.
Figure 38 shows cancer and cell-line vs. normal tissue expression.
Figure 39 shows cancer and cell-line vs. normal tissue expression.
Figure 40 shows cancer and cell-line vs. normal tissue expression.
Figure 41 shows cancer and cell-line vs. normal tissue expression.
Figure 42 shows cancer and cell-line vs. normal tissue expression.
Figure 43 is a histogram showing differential expression of a variety of transcripts in cancerous ovary samples relative to the normal samples.
Figure 44 shows cancer and cell-line vs. normal tissue expression.
DESCRIPTION OF PREFERRED EMBODIMENTS
The present invention is of novel markers for ovarian cancer that are both sensitive and accurate. Biomolecular sequences (amino acid and/or nucleic acid sequences) uncovered using the methodology of the present invention and described herein can be efficiently utilized as tissue or pathological markers and/or as drugs or drug targets for treating or preventing a disease.
Furthermore, at least certain of these markers are able to distinguish between various types of ovarian cancer, such as Ovarian epithelial tumors (serous, mucinous, endometroid, clear cell, and Brenner tumor), ovarian germ-cell tumors, (teratoma, dysgerminoma, endodermal sinus tumor, and embryonal carcinoma) and ovarian stromal tumors (originating from granulosa, theca, Sertoli, Leydig, and collagen-producing stromal cells), alone or in combination. These markers are differentially expressed, and preferably overexpressed in ovarian cancer specifically, as opposed to normal ovarian tissue. The measurement of these markers, alone or in combination, in patient samples provides information that the diagnostician can correlate with a probable diagnosis of ovarian cancer. The markers of the present invention, alone or in combination, show a high degree of differential detection between ovarian cancer and non-cancerous states.
The markers of the present invention, alone or in combination, can be used for prognosis, prediction, screening, early diagnosis, staging, therapy selection and treatment monitoring of ovarian cancer. For example, optionally and preferably, these markers may be used for staging ovarian cancer and/or monitoring the progression of the disease. Furthermore, the markers of the present invention, alone or in combination, can be used for detection of the source of metastasis found in anatomical places other thenovary. Also, one or more of the markers may optionally be used in combination with one or more other ovarian cancer markers (other than those described herein). According to an optional embodiment of the present invention, such a combination may be used to differentiate between various types of ovarian cancer, such as Ovarian epithelial tumors (serous, mucinous, endometroid, clear cell, and Brenner tumor), ovarian gerrrrcell tumors, (teratoma, dysgerminoma, endodermal sinus tumor, and embryonal carcinoma) and ovarian stromal tumors (originating from either granulosa, theca, Sertoli, Leydig, and collagerrproducing stromal cells).
These markers are specifically released to the bloodstream under conditions of ovarian cancer (or one of the above indicative conditions), and/or are otherwise expressed at a much higher level and/or specifically expressed in ovarian cancer tissue or cells, and/or tissue or cells under one of the above indicative conditions. The measurement of these markers, alone or in combination, in patient samples provides information that the diagnostician can correlate with a probable diagnosis of ovarian cancer and/or a condition that it is indicative of a higher risk for ovarian cancer.
The present invention therefore also relates to diagnostic assays for ovarian cancer, and methods of use of such markers for detection of ovarian cancer, optionally and preferably in a sample taken from a subject (patient), which is more preferably some type of blood sample.
In another embodiment, the present invention relates to bridges, tails, heads and/or insertions, and/or analogs, homologs and derivatives of such peptides. Such bridges, tails, heads and/or insertions are described in greater detail below with regard to the Examples.
As used herein a "tail" refers to a peptide sequence at the end of an amino acid sequence that is unique to a splice variant according to the present invention.
Therefore, a splice variant having such a tail may optionally be considered as a chimera, in that at least a first portion ofthe splice variant is typically highly homologous (often 100% identical) to a portion of the corresponding known protein, while at least a second portion of the variant comprises the tail.
As used herein a "head" refers to a peptide sequence at the beginning of an amino acid sequence that is unique to a splice variant according to the present invention. Therefore, a splice variant having such a head may optionally be considered as a chimera, in that at least a first portion of the splice variant comprises the head, while at least a second portion is typically highly homologous (often 100% identical) to a portion of the corresponding known protein.
As used herein "an edge portion" refers to a connection between two portions of a splice variant according to the present invention that were not joined in the wild type or known protein. An edge may optionally arise due to a join between the above "known protein" portion of a variant and the tail, for example, and/or may occur if an internal portion of the wild type sequence is no longer present, such that two portions of the sequence are now contiguous in the splice variant that were not contiguous in the known protein. A "bridge" may optionally be an edge portion as described above, but may also include a join between a head and a "known protein" portion of a variant, or a join between a tail and a "known protein"
portion of a variant, or a join between an insertion and a "known protein" portion of a variant.
Optionally and preferably, a bridge between a tail or a head or a unique insertion, and a "known protein" portion of a variant, comprises at least about 10 amino acids, more preferably at least about 20 amino acids, most preferably at least about 30 amino acids, and even more preferably at least about 40 amino acids, in which at least one amino acid is from the tail/head/insertion and at least one amino acid is from the "known protein"
portion of a variant.
Also optionally, the bridge may comprise any number of amino acids from about 10 to about 40 amino acids (for example, 10, 11, 12, 13...37, 38, 39, 40 amino acids in length, or any number in between).
It should be noted that a bridge cannot be extended beyond the length of the sequence in either direction, and it should be assumed that every bridge description is to be read in such manner that the bridge length does not extend beyond the sequence itself.
Furthermore, bridges are described with regard to a sliding window in certain contexts below. For example, certain descriptions of the bridges feature the following format: a bridge between two edges (in which a portion of the known protein is not present in the variant) may optionally be described as follows: a bridge portion of CONTIG-NAME PI
(representing the name of the protein), comprising a polypeptide having a length "n", wherein n is at least about amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most 10 preferably at least about 50 amino acids in length, wherein at least two amino acids comprise XX (2 amino acids in the center of the bridge, one from each end of the edge), having a structure as follows (numbering according to the sequence of CONTIG-NAME P I
): a sequence starting from any of amino acid numbers 49-x to 49 (for example); and ending at any of amino acid numbers 50 + ((n-2) - x) (for example), in which x varies from 0 to n-2.
In this example, it I 5 should also be read as including bridges in which n is any number of amino acids between 10-50 amino acids in length. Furthermore, the bridge polypeptide cannot extend beyond the sequence, so it should be read such that 49-x (for example) is not less than 1, nor 50 +
((r~2) - x) (for example) greater than the total sequence length.
In another embodiment, this invention provides antibodies specifically recognizing the splice variants and polypeptide fragments thereof of this invention.
Preferably such antibodies differentially recognize splice variants of the present invention but do not recognize a corresponding known protein (such known proteins are discussed with regard to their splice variants in the Examples below).
In another embodiment, this invention provides an isolated nucleic acid molecule encoding for a splice variant according to the present invention, having a nucleotide sequence as set forth in any one of the sequences listed herein, or a sequence complementary thereto. In another embodiment, this invention provides an isolated nucleic acid molecule, having a nucleotide sequence as set forth in any one of the sequences listed herein, or a sequence complementary thereto. In another embodiment, this invention provides an oligonucleotide of at least about 12 nucleotides, specifically hybridizable with the nucleic acid molecules of this invention. In another embodiment, this invention provides vectors, cells, liposomes and compositions comprising the isolated nucleic acids of this invention.
In another embodiment, this invention provides a method for detecting a splice variant according to the present invention in a biological sample, comprising:
contacting a biological sample with an antibody specifically recognizing a splice variant according to the present invention under conditions whereby the antibody specifically interacts with the splice variant in the biological sample but do not recognize known corresponding proteins (wherein the known protein is discussed with regard to its splice variants) in the Examples below), and detecting said interaction; wherein the presence of an interaction correlates with the presence of a splice variant in the biological sample.
In another embodiment, this invention provides a method for detecting a splice variant nucleic acid sequences in a biological sample, comprising: hybridizing the isolated nucleic acid molecules or oligonucleotide fragments of at least about a minimum length to a nucleic acid material of a biological sample and detecting a hybridization complex; wherein the presence of a hybridization complex correlates with the presence of a splice variant nucleic acid sequence in the biological sample.
According to the present invention, the splice variants described herein are norrlimiting examples of markers for diagnosing ovarian cancer. Each splice variant marker of the present invention can be used alone or in combination, for various uses, including but not limited to, prognosis, prediction, screening, early diagnosis, determination of progression, therapy selection and treatment monitoring of ovarian cancer.
According to optional but preferred embodiments of the present invention, any marker according to the present invention may optionally be used alone or combination. Such a combination may optionally comprise a plurality of markers described herein, optionally including any subcombination of markers, and/or a combination featuring at least one other marker, for example a known marker. Furthermore, such a combination may optionally and preferably be used as described above with regard to determining a ratio between a quantitative or semi-quantitative measurement of any marker described herein to any other marker described herein, and/or any other known marker, and/or any other marker. With regard to such a ratio between any marker described herein (or a combination thereof) and a known marker, more preferably the known marker comprises the "known protein" as described in greater detail below with regard to each cluster or gene.
According to other preferred embodiments of the present invention, a splice variant protein or a fragment thereof, or a splice variant nucleic acid sequence or a fragment thereof, may be featured as a biomarker for detecting ovarian cancer and/or an indicative condition, such that a biomarker may optionally comprise any of the above.
According to still other preferred embodiments, the present invention optionally and preferably encompasses any amino acid sequence or fragment thereof encoded by a nucleic acid sequence corresponding to a splice variant protein as described herein. Any oligopeptide or peptide relating to such an amino acid sequence or fragment thereof may optionally also (additionally or alternatively) be used as a biomarker, including but not limited to the unique amino acid sequences of these proteins that are depicted as tails, heads, insertions, edges or bridges. The present invention also optionally encompasses antibodies capable of recognizing, and/or being elicited by, such oligopeptides or peptides.
The present invention also optionally and preferably encompasses any nucleic acid sequence or fragment thereof, or amino acid sequence or fragment thereof, corresponding to a splice variant of the present invention as described above, optionally for any application.
Non-limiting examples of methods or assays are described below.
The present invention also relates to kits based upon such diagnostic methods or assays.
Nucleic acid sequences and Oligonucleotides Various embodiments of the present invention encompass nucleic acid sequences described hereinabove; fragments thereof, sequences hybridizable therewith, sequences homologous thereto, sequences encoding similar polypeptides with different codon usage, altered sequences characterized by mutations, such as deletion, insertion or substitution of one or more nucleotides, either naturally occurring or artificially induced, either randomly or in a targeted fashion.
The present invention encompasses nucleic acid sequences described herein;
fragments thereof, sequences hybridizable therewith, sequences homologous thereto [e.g., at least 50 %, at least 55 %, at least 60%, at least 65 %, at least 70 %, at least 75 %, at least 80 %, at least 85 %, at least 95 % or more say 100 % identical to the nucleic acid sequences set forth below], sequences encoding similar polypeptides with different codon usage, altered sequences characterized by mutations, such as deletion, insertion or substitution of one or more nucleotides, either naturally occurring or man induced, either randomly or in a targeted fashion. The present invention also encompasses homologous nucleic acid sequences (i.e., which form a part of a polynucleotide sequence of the present invention) which include sequence regions unique to the polynucleotides of the present invention.
In cases where the polynucleotide sequences of the present invention encode previously unidentified polypeptides, the present invention also encompasses novel polypeptides or portions thereof, which are encoded by the isolated polynucleotide and respective nucleic acid fragments thereof described hereinabove.
A "nucleic acid fragment" or an "oligonucleotide" or a "polynucleotide" are used herein interchangeably to refer to a polymer of nucleic acids. A polynucleotide sequence of the present invention refers to a single or double stranded nucleic acid sequences which is isolated and provided in the form of an RNA sequence, a complementary polynucleotide sequence (cDNA), a genomic polynucleotide sequence and/or a composite polynucleotide sequences (e.g., a combination of the above).
As used herein the phrase "complementary polynucleotide sequence" refers to a sequence, which results from reverse transcription of messenger RNA using a reverse transcriptase or any other RNA dependent DNA polymerase. Such a sequence can be subsequently amplified in vivo or in vitro using a DNA dependent DNA
polymerase.
As used herein the phrase "genomic polynucleotide sequence" refers to a sequence derived (isolated) from a chromosome and thus it represents a contiguous portion of a chromosome.
As used herein the phrase "composite polynucleotide sequence" refers to a sequence, which is composed of genomic and cDNA sequences. A composite sequence can include some exonal sequences required to encode the polypeptide of the present invention, as well as some intronic sequences interposing therebetween. The intronic sequences can be of any source, including of other genes, and typically will include conserved splicing signal sequences. Such intronic sequences may further include cis acting expression regulatory elements.
Preferred embodiments of the present invention encompass oligonucleotide probes.
An example of an oligonucleotide probe which can be utilized by the present invention is a single stranded polynucleotide which includes a sequence complementary to the unique sequence region of any variant according to the present invention, including but not limited to a nucleotide sequence coding for an amino sequence of a bridge, tail, head and/or insertion according to the present invention, and/or the equivalent portions of any nucleotide sequence given herein (including but not limited to a nucleotide sequence of a node, segment or amplicon described herein).
Alternatively, an oligonucleotide probe of the present invention can be designed to hybridize with a nucleic acid sequence encompassed by any of the above nucleic acid sequences, particularly the portions specified above, including but not limited to a nucleotide sequence coding for an amino sequence of a bridge, tail, head and/or insertion according to the present invention, and/or the equivalent portions of any nucleotide sequence given herein (including but not limited to a nucleotide sequence of a node, segment or amplicon described herein).
Oligonucleotides designed according to the teachings of the present invention can be generated according to any oligonucleotide synthesis method known in the art such as enzymatic synthesis or solid phase synthesis. Equipment and reagents for executing solid-phase synthesis are commercially available from, for example, Applied Biosystems. Any other means for such synthesis may also be employed; the actual synthesis of the oligonucleotides is well within the capabilities of one skilled in the art and can be accomplished via established methodologies as detailed in, for example, "Molecular Cloning: A laboratory Manual" Sambrook et al., (1989);
"Current Protocols in Molecular Biology" Volumes I-III Ausubel, R. M., ed.
(1994); Ausubel et al., "Current Protocols in Molecular Biology", John Wiley and Sons, Baltimore, Maryland (1989); Perbal, "A Practical Guide to Molecular Cloning", John Wiley & Sons, New York (1988) and "Oligonucleotide Synthesis" Gait, M. J., ed. (1984) utilizing solid phase chemistry, e.g. cyanoethyl phosphoramidite followed by deprotection, desalting and purification by for example, an automated trityl-on method or HPLC.
Oligonucleotides used according to this aspect of the present invention are those having a length selected from a range of about 10 to about 200 bases preferably about 15 to about 150 bases, more preferably about 20 to about 100 bases, most preferably about 20 to about 50 bases.
Preferably, the oligonucleotide of the present invention features at least 17, at least 18, at least 19, at least 20, at least 22, at least 25, at least 30 or at least 40, bases specifically hybridizable with the biomarkers of the present invention.
The oligonucleotides of the present invention may comprise heterocylic nucleosides consisting ofpurines and the pyrimidines bases, bonded in a 3' to 5' phosphodiester linkage.
Preferably used oligonucleotides are those modified at one or more of the backbone, internucleoside linkages or bases, as is broadly described hereinunder.
Specific examples of preferred oligonucleotides useful according to this aspect of the present invention include oligonucleotides containing modified backbones or norrnatural internucleoside linkages. Oligonucleotides having modified backbones include those that retain a phosphorus atom iri the backbone, as disclosed in U.S. Pat. NOs: 4,469,863;
4,476,301;
5,023,243; 5,177,196; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717;
5,321,131;
5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466, 677; 5,476,925; 5,519,126;
5,536,821;
5,541,306; 5,550,111; 5,563,253; 5,571,799; 5,587,361; and 5,625,050.
Preferred modified oligonucleotide backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkyl phosphotriesters, methyl and other alkyl phosphonates including 3'-alkylene phosplnnates and chiral phosphonates, phosphinates, phosphoramidates including 3'-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates having normal 3'-5' linkages, 2'-5' linked analogs of these, and those having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'. Various salts, mixed salts and free acid forms can also be used.
Alternatively, modified oligonucleotide backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatom and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside);
siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones;
methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S and CHI component parts, as disclosed in U.S. Pat. Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134;
5,216,141; 5,235,033;
5,264,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; 5,489,677;
5,541,307;
5,561,225; 5,596,086; 5,602,240; 5,610,289; 5,602,240; 5,608,046; 5,610,289;
5,618,704; 5,623, 070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439.
Other oligonucleotides which can be used according to the present invention, are those modified in both sugar and the internucleoside linkage, i.e., the backbone, of the nucleotide units are replaced with novel groups. The base units are maintained for complementation with the appropriate polynucleotide target. An example for such an oligonucleotide mimetic, includes peptide nucleic acid (PNA). United States patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, each of which is herein incorporated by reference. Other backbone modifications, which can be used in the present invention are disclosed in U.S. Pat. No: 6,303,374.
Oligonucleotides of the present invention may also include base modifications or substitutions. As used herein, "unmodified" or "natural" bases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U).
Modified bases include but are not limited to other synthetic and natural bases such as S-methylcytosine (5-me-C), S~hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly S-bromo, ~trifluoromethyl and other S-substituted uracils and cytosines, '~methylguanine and '~methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine.
Further bases particularly useful for increasing the binding affinity of the oligomeric compounds of the invention include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, including 2 aminopropyladenine, S~propynyluracil and ~propynylcytosine.
5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6 1.2 °C and are presently preferred base substitutions, even more particularly when combined with 2'-O-methoxyethyl sugar modifications.
Another modification of the oligonucleotides of the invention involves chemically linking to the oligonucleotide one or more moieties or conjugates, which enhance the activity, cellular distribution or cellular uptake of the oligonucleotide. Such moieties include but are not limited to lipid moieties such as a cholesterol moiety, cholic acid, a thioether, e.g., hexyl-S-tritylthiol, a thiocholesterol, an aliphatic chain, e.g., dodecandiol or undecyl residues, a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate, a polyamine or a polyethylene glycol chain, or adamantine acetic acid, a palmityl moiety, or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety, as disclosed in U.S. Pat. No: 6,303,374.
It is not necessary for all positions in a given oligonucleotide molecule to be uniformly modified, and in fact more than one of the aforement Toned modifications may be incorporated in a single compound or even at a single nucleoside within an oligonucleotide.
It will be appreciated that oligonucleotides of the present invention may include further modifications for more efficient use as diagnostic agents and/or to increase bioavailability, therapeutic efficacy and reduce cytotoxicity.
To enable cellular expression of the polynucleotides of the present invention, a nucleic acid construct according to the present invention may be used, which includes at least a coding region of one of the above nucleic acid sequences, and further includes at least one cis acting regulatory element. As used herein, the phrase "cis acting regulatory element"
refers to a polynucleotide sequence, preferably a promoter, which binds a trans acting regulator and regulates the transcription of a coding sequence located downstream thereto.
Any suitable promoter sequence can be used by the nucleic acid construct of the present invention.
Preferably, the promoter utilized by the nucleic acid construct of the present invention is active in the specific cell population transformed. Examples of cell type-specific and/or tissue-specific promoters include promoters such as albumin that is liver specific, lymphoid specific promoters [Calame et al., (1988) Adv. Immunol. 43:235-275]; in particular promoters of T cell receptors [Winoto et al., (1989) EMBO J. 8:729-733] and immunoglobulins;
[Banerji et al.
(1983) Cell 33729-740], neurorrspeciflc promoters such as the neurofilament promoter [Byrne et al. (1989) Proc. Natl. Acid. Sci. USA 86:5473-5477], pancreas-specific promoters [Edlunch et al. (1985) Science 230:912-916] or mammary gland-specific promoters such as the milk whey promoter (U.S. Pat. No. 4,873,316 and European Application Publication No. 264,166).
The nucleic acid construct of the present invention can further include an enhancer, which can be adjacent or distant to the promoter sequence and can function in up regulating the transcription therefrom.
The nucleic acid construct of the present invention preferably further includes an appropriate selectable marker and/or an origin of replication. Preferably, the nucleic acid construct utilized is a shuttle vector, which can propagate both in E. coli (wherein the construct comprises an appropriate selectable marker and origin of replication) and be compatible for propagation in cells, or integration in a gene and a tissue of choice. The construct according to the present invention can be, for example, a plasmid, a bacmid, a phagemid, a cosmid, a phage, a virus or an artificial chromosome.
Examples of suitable constructs include, but are not limited to, pcDNA3, pcDNA3.1 (+/-), pGL3, PzeoSV2 (+/-), pDisplay, pEF/myc/cyto, pCMV/myc/cyto each of which is commercially available from Invitrogen Co. (www.invitrogen.com). Examples of retroviral I S vector and packaging systems are those sold by Clontech, San Diego, Calif., includingRetro-X
vectors pLNCX and pLXSN, which permit cloning into multiple cloning sites and the trasgene is transcribed from CMV promoter. Vectors derived from Mo-MuLV are also included such as pBabe, where the transgene will be transcribed from the 5'LTR promoter.
Currently preferred in vivo nucleic acid transfer techniques include transfection with viral or norrviral constructs, such as adenovirus, lentivirus, Herpes simplex I virus, or adeno associated virus (AAV) and lipid-based systems. Useful lipids for lipid-mediated transfer of the gene are, for example, DOTMA, DOPE, and DC-Chol [Tonkinson et al., Cancer Investigation, 14(1): 54-65 (1996)]. The most preferred constructs for use in gene therapy are viruses, most preferably adenoviruses, AAV, lentiviruses, or retroviruses. A viral construct such as a retroviral construct includes at least one transcriptional promoter/enhancer or locus-defining element(s), or other elements that control gene expression by other means such as alternate splicing, nuclear RNA export, or post-translational modification of messenger.
Such vector constructs also include a packaging signal, long terminal repeats (LTRs) or portions thereof, and positive and negative strand primer binding sites appropriate to the virus used, unless it is already present in the viral construct. In addition, such a construct typically includes a signal sequence for secretion of the peptide from a host cell in which it is placed.
Preferably the signal sequence for this purpose is a mammalian signal sequence or the signal sequence of the polypeptide variants of the present invention. Optionally, the construct may also include a signal that directs polyadenylation, as well as one or more restriction sites and a translation termination sequence. By way of example, such constructs will typically include a 5' LTR, a tRNA binding site, a packaging signal, an origin of second-strand DNA
synthesis, and a 3' LTR
or a portion thereof. Other vectors can be used that are norrviral, such as cationic lipids, polylysine, and dendrimers.
Hybridization assays Detection of a nucleic acid of interest in a biological sample may optionally be effected by hybridizatior~based assays using an oligonucleotide probe (non-limiting examples of probes according to the present invention were previously described).
Traditional hybridization assays include PCR, RT-PCR, Real-time PCR, RNase protection, in-situ hybridization, primer extension, Southern blots (DNA
detection), dot or slot blots (DNA, RNA), and Northern blots (RNA detection) (NAT type assays are described in greater detail below). More recently, PNAs have been described (Nielsen et al.
1999, Current Opin. Biotechnol. 10:71-75). Other detection methods include kits containing probes on a dipstick setup and the like.
Hybridization based assays which allow the detection of a variant of interest (i.e., DNA
or RNA) in a biobgical sample rely on the use of oligonucleotides which can be 10, 15, 20, or to 100 nucleotides long preferably from 10 to 50, more preferably from 40 to 50 nucleotides long.
Thus, the isolated polynucleotides (oligonucleotides) of the present invention are preferably hybridizable with any of the herein described nucleic acid sequences under moderate 25 to stringent hybridization conditions.
Moderate to stringent hybridization conditions are characterized by a hybridization solution such as containing 10 % dextrane sulfate, 1 M NaCI, 1 % SDS and 5 x 106 cpm 32P
labeled probe, at 65 °C, with a final wash solution of 0.2 x SSC and 0.1 % SDS and final wash at 65°C and whereas moderate hybridization is effected using a hybridization solution 30 containing 10 % dextrane sulfate, 1 M NaCI, 1 % SDS and 5 x 106 cpm 32P
labeled probe, at 65 °C, with a final wash solution of 1 x SSC and 0.1 % SDS and final wash at SO °C.
More generally, hybridization of short nucleic acids (below 200 by in length, e.g. 17-40 by in length) can be effected using the following exemplary hybridization protocols which can be modified according to the desired stringency; (i) hybridization solution of 6 x SSC and 1 SDS or 3 M TMACI, 0.01 M sodium phosphate (pH 6.8), 1 mM EDTA (pH 7.6), 0.5 %
SDS, 100 pg/ml denatured salmon sperm DNA and 0.1 % nonfat dried milk, hybridization temperature of 1 - 1.5 °C below the TIn, final wash solution of 3 M TMACI, 0.01 M
sodium phosphate (pH
6.8), 1 mM EDTA (pH 7.6), 0.5 % SDS at 1 - 1.5 °C below the Tm; (ii) hybridization solution of 6 x SSC and 0.1 % SDS or 3 M TMACI, 0.01 M sodium phosphate (pl-I 6.8), 1 mM EDTA
(pH 7.6), 0.5 % SDS, 100 pg/ml denatured salmon sperm DNA and 0.1 % nonfat dried milk, hybridization temperature of 2 - 2.5 °C below the Tm, final wash solution of 3 M TMACI, 0.01 M sodium phosphate (pH 6.8), 1 mM EDTA (pH 7.6), 0.5 % SDS at 1 - 1.5 °C below the Tm, final wash solution of 6 x SSC, and final wash at 22 °C; (iii) hybridization solution of 6 x SSC
and 1 % SDS or 3 M TMACI, 0.01 M sodium phosphate (pH 6.8), 1 mM EDTA (pH
~ ~~M =
tem N ~~ .~
iie~
o ai s r~' ~ ~
~ ~. w ~~ ~ .~
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~TranSCript Name -.
~3.r ~TA.
Yk ~.
~"
PEA
a nucleic acid sequence comprising a sequence in the table below:
"ms's ~x~~~"''~' a'""~"F
' ~x Segment~~~lame ~~-~ _ ~ ~",.
M
z"
i"' , ~"yet, .F
HSCP2 PEA node 0 HSCP2 PEA node 3 HSCP2 PEA node 6 HSCP2 PEA node 8 HSCP2 PEA node 10 HSCP2 PEA node 14 HSCP2 PEA node 23 HSCP2 PEA node 26 HSCP2 PEA node 29 HSCP2 PEA node 31 HSCP2 PEA node 32 I
HSCP2_PEA node 34 HSCP2 I node 52 ' PEA
HSCP2 1 node 58 PEA
HSCP2 1 node 72 PEA
HSCP2 1 node 73 PEA
HSCP2 1 node 74 PEA
HSCP2 1 node 76 PEA
HSCP2 1 node 78 PEA
HSCP2 1 node 80 PEA
HSCP2 1 node 84 PEA
HSCP2 1 node 4 PEA
HSCP2 1 node 7 PEA
HSCP2 1 node 13 PEA
HSCP2 1 node I S
PEA
HSCP2 1 node 16 PEA
HSCP2 1 node 18 PEA
HSCP2 1 node 20 PEA
HSCP2 1 node 21 PEA
HSCP2 1 node 37 PEA
HSCP2 1 node 38 PEA
HSCP2 1 node 39 PEA
HSCP2 1 node 41 PEA
HSCP2 1 node 42 PEA
HSCP2 1 node 46 PEA
HSCP2 1 node 47 PEA
HSCP2 1 node SO
PEA
HSCP2 1 node 51 PEA
HSCP2 1 node 55 PEA
HSCP2 1 node 56 PEA
HSCP2 1 node 60 PEA
HSCP2 1 node 61 PEA
HSCP2 PEA node67 HSCP2 PEA node68 HSCP2 PEA node69 HSCP2 PEA node70 I-ISCP2 node75 HSCP2 PEA node77 I
HSCP2 PEA node79 HSCP2 PEA node82 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
"'~"'.r'4.'~";~i3 ~ ..a~~
~,!'.4 ~ '~ &
, S ~.
.~
_,~ k~l I k ,. S. , ~r 'i;.",&
-~' ,.~,~;.
t~. ., ~,:,'~~
~. 3i' ;,~ ..,,y>'a:"'e Prorexn Name _ ~ , ~ ~
_ ~ ~~;, ~ .,~ :v~
~ .,; .
"..,...g~
w'~~~.
" x E:
fi~ s ..~-y~.x'~
~'e~ fli, '.W>p _ h~S_ ~' "G ;i~5 A~ ~ ,s , ,t'...
v s ~ ~T..
-' e. ' A....i"~
~ P'~..
.
.~2s~~~
~:,...~:aa ...._:.-zur:
'a-f .,~',.e"a'-' ': ;
' y ~ ~
- . _ k~.
., ~ -n~.
..a -- ~,..ma....e.
~~ ~ ,.,~
r.....,'?~~
.~Tlz _ .
I P4 s I
I
I
I
I
I
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
E ~~ ~ ~~ ~H
Traos tlpt Nam e~
t r~
HUMTEN~~PEA I T4 ( I
I
PEA
PEA
PEA
PEA
a nucleic acid sequence comprising a sequence in the table below:
~~ m~~~
~' ~~ ,~~
~a~ ~ ~,;
S~ e~t~Narne ~: _ T
HUMTEN PEA 1 node 0 HUMTEN PEA 1 node 2 HUMTEN PEA 1 node 5 HUMTEN PEA I node 6 HUMTEN PEA 1 node 11 HUMTEN PEA 1 node 12 HUMTEN PEA 1 node 16 HUMTEN PEA 1 node t9 HUMTEN PEA 1 node 23 HUMTEN PEA 1 node 27 I-IUMTEN 1 node 28 PEA
HUMTEN PEA 1 node 30 HUMTEN PEA 1 node 32 HUMTEN PEA I node 33 I-IUMTEN 1 node 35 PEA
1-IUMTEN l node 38 PEA
HUMTEN PEA 1 node 40 HUMTEN PEA 1 node 42 HUMTEN PEA 1 node 43 HUMTEN PEA 1 node 44 HUMTEN PEA 1 node 45 HUMTEN .PEA1 node 46 HUMTEN PEA 1 node 47 HUMTEN PEA 1 node 49 HUMTEN PEA 1 node 51 HUMTEN PEA 1 node 56 HUMTEN PEA 1 node 65 HUMTEN PEA 1 node 71 HUMTEN PEA 1 node 73 HUMTEN PEA 1 node 76 HUMTEN PEA 1 node 79 HUMTEN PEA 1 node 83 HUMTEN PEA 1 node 89 HUMTEN PEA 1 node 7 HUMTEN PEA 1 node 8 HUMTEN PEA 1 node 9 HUMTEN PEA_1 node_14 HUMTEN PEA 1 node 17 HUMTEN PEA 1 node 21 HUMTEN PEA 1 node 22 HUMTEN PEA 1 node 25 HUMTEN PEA 1 node 36 HUMTEN PEA 1 node 53 HUMTEN PEA 1 node 54 HUMTEN PEA 1 node 57 HUMTEN PEA 1 node 61 HUMTEN PEA 1 node 62 HUMTEN PEA 1 node 67 HUMTEN PEA 1 node 68 HUMTEN PEA 1 node 69 HUMTEN PEA 1 node 70 HUMTEN PEA 1 node 72 HUMTEN PEA 1 node 84 HUMTEN PEA 1 node 85 HUMTEN PEA 1 node 86 HUMTEN PEA 1 node 87 HUMTEN PEA 1 node 88 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
'~"~ a ~~
aid'. ~r ~','t' '~' .'~ ~:-r ~. "
otean ame .. ",: , ~~.w,.~,sr~x"~.4~~--.';~','~;~~2.x:..:~~.._s~-,r~'-~:.,.~;_a":~,~'~'~z..~~b4;,i .5":,h;w:~._.:
HUMTEN PEA PS
I
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~r~'Y"~' ~
;~ .,~ z 2.~Y. .,.
,~~~,,, .
r r~ s ~ . 3s ~T~an~~'rlpt ~'G~~
.' a , . .~
r ~
..
~~
PEA
PEA
a nucleic acid sequence comprising a sequence in the table below:
a, egme~t ~,arn~ ~ ,~ 'r 'c-x ~~c s~
HUMOSTRO PEA 1~ PEA 1 node 0 HUMOSTRO PEA 1 PEA 1 node 10 HUMOSTRO PEA 1 PEA 1 node 16 HUMOSTRO PEA 1 PEA 1 node 23 1-IUMOSTRO 1 PEA node31 I-1UMOSTRO 1 PEA node HUMOSTRO PEA 1 PEA node3 HUMOSTRO PEA 1 PEA node5 HUMOSTRO PEA I PEA node7 HUMOSTRO PEA I PEA node8 HUMOSTRO PEA 1 PEA nodeI S
HUMOSTRO PEA 1 PEA node17 HUMOSTRO PEA I PEA node20 HUMOSTRO PEA 1 PEA node21 HUMOSTRO PEA 1 PEA node22 HUMOSTRO PEA 1 PEA node24 I
HUMOSTRO PEA 1 PEA node26 HUMOSTRO PEA 1 PEA node27 HUMOSTRO PEA I PEA node HUMOSTRO PEA I PEA node HUMOSTRO PEA 1 PEA node30 I
HUMOSTRO PEA I PEA node32 I
HUMOSTRO PEA I PEA node34 HUMOSTRO PEA 1 PEA node36 HUMOSTRO PEA 1 PEA node37 HUMOSTRO PEA I PEA node38 I
HUMOSTRO PEA 1 PEA node39 HUMOSTRO PEA 1 PEA node HUMOSTRO PEA I PEA node HUMOSTRO PEA I PEA node According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~_. ,~
-~:~....> .
':~~ ~ ~, ;.-y -: i ;r .
P otyn Name ,.~.a ~ ;. ~-~ ~. ~,...
.~ ~~ , d ~ . ~
3 .. ~ >!.
~' ,. ~
z '.
~
'.' ~
~...~
.. , a dt :
..,:--8,1 .. d._ y. .~.,0.~"-w '..:~'F.- .: ~~~... ~
,. ;r...kw ~~, .x,a4~& ,.,:~ , ~s,_..
t ~
~
~
HUMOSTRO -, PEA 1 .
PEA
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~Transcnpt Nam '~ ~ ~
~ ~~~':ar"..
f.~ 'k:.~
.. r.~sW
.e~ .CEO
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA_1 a nucleic acid sequence comprising a sequence in the table below:
~~SegrnentName a~l~ ,r'' ~ "~ ~r .'..
_..,:
T46984 PEA node 2 I
T46984 PEA node 4 T46984 PEA node 6 I
T46984 PEA node 12 T46984 PEA node 14 T46984 PEA node 25 T46984 PEA node 29 I
T46984 PEA node 34 I
T46984 PEA node 46 T46984 PEA node 47 I
T46984 PEA node 52 T46984 PEA node 65 I
T46984 PEA node 69 T46984 PEA node 75 T46984 PEA node 86 I
T46984 PEA node 9 T46984 PEA node 13 T46984 PEA node 19 T46984 PEA node 21 T46984 PEA node 22 T46984 PEA node 26 T46984 PEA node 28 node node node node node node node node node node node node node node node node node node node node node node node node node node ~T46984PEA-1 85 node According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
_. ~,. ~
s~. 't y ~ - Y.~
~~ ~_, ~~ =
rote~n Name y~ w. ' ~ , T~.~
~ ..~.~
A~. w ~
.'~f. y'~a.
..~ y~
~ .~~..wT
.~, i,, , R " ~i.yofi~...
'.:
"i- ~ ~
' ''F.'~
'T'L.-Y~'m . ~. x.~.w~'~i' (. ~y '-:
'z'~f Y:
..., _4~.
~,.ra.~
,.,:.e:
A., t.'3dd~-'s-..";".'ki ~ r= ."
~ m '~'~'.~..'~,~,~
2., .t .' I;~."
x' 3..
~ - ~a5 '~,;'v s.~;,.~~-s~aC
, ,.:..A;',#Y;R
I
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
Transcrap Nine ~'p ~X~~
~f~~~~~
' :,~ ' a nucleic acid sequence comprising a sequence in the table below:
~Se '~ dam ~ ~° ~ y;
a , ~ " ~ .r ~. . ~' 'y M78530 PEA 1 node 0 M78530 PEA 1 node 15 M78530 PEA 1 node 16 M78530 PEA-1 node 19 node node node node node node node node node node node node node node node node M78530PEA_1 34 node According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below andlor:
ranscName ~'~~
r '__ a nucleic acid sequence comprising a sequence in the table below:
Segment Name '' ~ ~.~
,: ~ ;:
_".. w_...__~ _. ~ . _.
T48119 node 0 T48119 node I1 T48119 node 13 T48119 node 38 T48119 node 41 T48119 node 45 T48119 node 47 T48119 node 4 T48119 node 8 T48119 node 15 T48119 node 17 T48119 node 20 T48119 node 22 T48119 node 26 T48119 node 28 T48119 node 31 T48119 node 32 T48119 node 33 T48119 node 44 According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~~Pro "~am~ ~ ~ ~ °- ~ ~ ~' ~~ ~ ~ ,err.
According to preferred embodiments of the present invention, there is provided an isolated polynucleotide comprising a nucleic acid sequence in the table below and/or:
~TransCriptName~
~~
-~: ~~,~~' %.~. _ ~ ~:
_ .~n~ t $
I
I
I
I
~HSMUC 1 A T47 PEA_ 1 a nucleic acid sequence comprising a sequence in the table below:
Se~gniien~~
P~ 3 ~ ;.
x ~''~ ~~
~ m HSMUC 1 A PEA node 0 HSMUC 1 A PEA node 14 HSMUC 1 A PEA node 24 HSMUC I A PEA node 29 HSMUC1A PEA node 35 HSMUCIA PEA node 38 HSMUC1A PEA_1 node 3 A PEA node PEA node PEA node PEA node node node HSMUC1A PEA l 20 node node node node node node PEA node node node According to preferred embodiments of the present invention, there is provided an isolated polypeptide comprising an amino acid sequence in the table below:
~~a~t~~~e "',a. - 't', ~,, d ~s<r, -e..-..e. .,'~.
a, ~M '-'~'3,~e' a ;. ~~g~~Z
,r~ ", ~
~~s.~, ~~ - ~ "~, m ~'.n~' ~;.z,a.R,~
'~._~ _._.'~_a..m,.
>~a PEA
PEA
PEA
PEA
PEA
PEA
PEA
PEA
~SMUC 1 A 1 P52 PEA-f-1SMUC1APEA I P53 I A
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSMUCIA PEA-1 P63, comprising a first amino acid sequence being at least 90 % homologous to MTPGTQSPFFLLLLLTVLTVVTGSGHASSTPGGEKETSATQRSSV corresponding to amino acids I - 45 of MUC1 HUMAN, which also corresponds to amino acids 1 - 45 of HSMUCIA PEA-1 P63, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence EEEVSADQVSVGASGVLGSFKEARNAPSFLSWSFSMGPSK corresponding to amino acids 46 - 85 of HSMUC I A PEA-1 P63, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSMUCIA PEA_1 P63, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence EEEVSADQVSVGASGVLGSFKEARNAPSFLSWSFSMGPSK in HSMUCIA PEA 1 P63.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA 1 P2, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGWLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVWVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDN
RYIANTVELRVKISTEVGITNVDLSTVDKDQSIAPKTTRVTYPAKAKGTFIADSHQNFAL
FFQLV DVNTGAELTPHQTFVRLHNQKTGQEV VFVAEPDNKNVYKFELDTSERKlEFDS
ASGTYTLYLI1GDATLKNPILWNV corresponding to amino acids 1 - 498 of RIB2 HUMAN, which also corresponds to amino acids 1 - 498 of T46984 PEA-1 P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VCA corresponding to amino acids 499 - 501 of T46984 PEA_1 P2, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P3, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPV V VVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTP VGDVFELNFMNVKFSSGYYDFLVEVEGDN
RYIANTVELRVKISTEVGITNVDLSTVDKDQSIAPKTTRVTYPAKAKGTFIADSHQNFAL
FFQLVDVNTGAELTPHQ corresponding to amino acids 1 - 433 of RIB2 HUMAN, which also corresponds to amino acids 1 - 433 of T46984 PEA-1 P3, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence ICHIWKLIFLP corresponding to amino acids 434 - 444 of T46984 PEA-1 P3, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA 1 P3, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence ICHIWKLIFLP in T46984 PEA 1 P3.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA_1 P10, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKI-IDVERLKASLDRPFTNLESAFYSIVGLSSL
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDN
FFQLVDVNTGAELTPHQTFVRLHNQKTGQEVVFVAEPDNKNVYKFELDTSERKIEFDS
ASGTYTLYLIIGDATLKNPILWNV corresponding to amino acids 1 - 498 of RIB2 HUMAN, which also corresponds to amino acids 1 - 498 of T46984 PEA 1 P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more 15 preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence LMDQK corresponding to amino acids 499 - 503 of T46984 PEA_1 P10, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an 20 isolated polypeptide encoding for a tail of T46984 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homolo gous to the sequence LMDQK
in T46984 PEA 1 P10.
According to preferred embodiments of the present invention, there is provided an 25 isolated chimeric polypeptide encoding for T46984 PEA_1 P11, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDN
RYIANTVELRVKISTEVGITN V DLSTVDKDQSIAPKTTRVTYPAKAKGTFIADSHQNFAL
FFQLVDVNTGAELTPHQTFV RLHINQKTGQEVVFVAEPDNKNVYKFELDTSERKIEFDS
ASGTYTLYLIIGDATLKNPILWNVADV VIKFPEEEAPSTVLSQNLFTPKQEIQHLFREPEK
RPPTVVSNTFTALILSPLLLLFALWIRIGANVSNFTFAPSTIIFHLGHAAMLGLMYVYWT
QLNMFQTLKYLAILGSVTFLAGNRMLAQQAVKR corresponding to amino acids 1 - 628 of RIB2 HUMAN, which also corresponds to amino acids 1 - 628 of T46984 PEA 1 PI
1.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984_PEA_1-P12, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAF YSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVGDVFELNFMN corresponding to amino acids 1 - 338 of RIB2_HUMAN, which also corresponds to amino acids 1 - 338 of T46984 PEA_1 P12, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence SQDLH corresponding to amino acids 339 -343 of T46984 PEA_1 P12, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA-1 P12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SQDLH in T46984 PEA 1 P 12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA_1 P21, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence M
corresponding to amino acids 1 - I of T46984 PEA-l P21, and a second amino acid sequence being at least 90 % homologous to KACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSSVTQIYHAV
AALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSIVEEIEDLVA
RLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNAIFSKKNFES
LSEAFSVASAAAVLSHNRYH V PV V V VPEGSASDTHEQAILRLQVTN VLSQPLTQATVKL
EHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDNRYIANTVEL
RVKISTEVGITNVDLSTVDKDQSIAPKTTRVTYPAKAKGTFIADSHQNFALFFQLVDVNT
GAELTPHQTFVRLHNQKTGQEVVFVAEPDNKNVYKFELDTSERKIEFDSASGTYTLYLII
GDATLKNPILWNVADVVIKFPEEEAPSTVLSQNLFTPKQEIQHLFREPEKRPPTVVSNTF
TALILSPLLLLFALWIRIGANVSNFTFAPSTIIFHLGHAAMLGLMYVYWTQLNMFQTLKY
LAILGSVTFLAGNRMLAQQAVKRTAH corresponding to amino acids 70 - 631 of RIB2 HUMAN, which also corresponds to amino acids 2 - 563 of T46984 PEA-1 P21, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA 1 P27, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKL,EHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDN
RYIANTVELRVKISTEVGITNVDLSTVDKDQSIAPKTTRVTYPAKAKGTFIADSHQNFA
corresponding to amino acids 1 - 415 of RIB2 HUMAN, which also corresponds to amino acids 1 - 415 of T46984 PEA-1 P27, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence FGSGLVPMSPTSLLLLARLYFTWDMLLCWDSCMSTGLSSTCSRP corresponding to amino acids 416 - 459 of T46984 PEA_1 P27, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA_1 P27, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence FGSGLVPMSPTSLLLLARLYFTWDMLLCWDSCMSTGLSSTCSRP in T46984 PEA 1 P27.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P32, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKI-IDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVGDVFELNFMNVKFSSGYYDFLVEVEGDN
RYIANTVE corresponding to amino acids 1 - 364 of RIB2 HUMAN, which also corresponds to amino acids 1 - 364 of T46984 PEA_1 P32, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GQVRWLTPVIPALWEAKAGGSPEVRSSILAWPT corresponding to amino acids 365 - 397 of T46984 PEA_1 P32, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA-1 P32, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GQVRWLTPVIPALWEAKAGGSPEVRSSILAWPT in T46984 PEA 1 P32.
According to preferred embodiments of the preset invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P34, comprising a first amino acid sequence being at least 90'% homologous to VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAILRLQVTNVLSQ
PLTQATVKLEHAKSVASRATVLQKTSFTPVG corresponding to amino acids 1 - 329 of RIB2 HUMAN, which also corresponds to amino acids 1 - 329 of T46984 PEA 1 P34.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P35, compris ing a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLATVQALQTASHLSQQADLRSI
VEEIEDLVARLDELGGVYLQFEEGLETTALFVAATYKLMDHVGTEPSIKEDQVIQLMNA
IFSKKNFESLSEAFSVASAAAVLSHNRYHVPVVVVPEGSASDTHEQAI corresponding to amino acids 1 - 287 of RIB2 HUMAN, which also corresponds to amino acids 1 -287 of T46984 PEA-1 P35, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 9S%
homologous to a polypeptide having the sequence GCWPSRQSREQHISSRRKMEILKTECQEKESRTIHSMRRKMEKKNFI corresponding to amino acids 288 - 334 of T46984 PEA_1 P35, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA-1 P35, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GCWPSRQSREQHISSRRKMEILKTECQEKESRTIHSMRRKMEKKNFI in T46984 PEA 1 P35.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T469$4 PEA-1 P38, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKI-iDVERLKASLDRPFTNLESAFYSI VGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCEISISNETKDLLLAAVSEDSS
VTQIYHAVAALSGFGLPLASQEAL corresponding to amino acids 1 - 145 of 5 R1B2 HUMAN, which also corresponds to amino acids 1 - 145 of T46984 PEA-1 P38, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MDPDWCQCLQLHFCS corresponding to amino acids 146 - 160 of T46984 PEA-1 P38; wherein said first amino acid sequence and second amino acid sequence 10 are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA_1 P38, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence 15 MDPDWCQCLQLHFCS in T46984 PEA 1 P38.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P39, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
VTQIYHAVAALSGFGLPLASQEALSALTARLSKEETVLA corresponding to amino acids 1 160 of RIB2 HUMAN, which also corresponds to amino acids 1 - 160 of T46984 PEA
1 P39.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P45, comprising a first amino acid 25 sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLTKHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAKKACTYIRSNLDPSNVDSLFYAAQASQALSGCE corresponding to amino acids 1 - 101 of RIB2 HUMAN, which also corresponds to amino acids 1 - 101 of T46984 PEA-1 P45, and a second amino acid sequence being at least 70%, optionally at least 30 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence NSPGSADSIPPVPAG corresponding to amino acids 102 - 1 16 of T46984 PEA_I P45, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984~PEA-1 P45, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NSPGSADSIP.PVPAG in T46984 PEA 1 P45.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T46984 PEA-1 P46, comprising a first amino acid sequence being at least 90 % homologous to MAPPGSSTVFLLALTIIASTWALTPTHYLT.KHDVERLKASLDRPFTNLESAFYSIVGLSSL
GAQVPDAK corresponding to amino acids 1 - 69 of RIB2,HUMAN, which also corresponds to amino acids 1 - 69 of T46984 PEA I P46, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NSPGSADSIPPVPAG corresponding to amino acids 70 - 84 of T46984~PEA-1 P46, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T46984 PEA-1 P46, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NSPGSADSIPPVPAG in T46984 PEA 1 P46.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA-1 P15, comprising a first amino acid sequence being at least 90 % homologous to MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEE
ETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKL
CEQDSTFDGVTDKPILDCCACGTAKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSH
SKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLN
DAGTDSGVTYESPNKPTIPQEKIRPLTSLDI-IPQSPFYDPEGGSITQVARVVIERIARKGEQ
CNIVPDNVDDIVADLAPEEKDEDDTPETCIYSNWSPWSACSSSTCDKGKRMRQRMLKA
QLDLSVPCPDTQDFQPCMGPGCSDEDGSTCTMSEWITWSPCSISCGMGMRSRERYVKQ
FPEDGSVCTLPTEE corresponding to amino acids 1 - 544 of Q9HCB6, which also corresponds to amino acids 1 - 544 of M78530 PEA_1 P15, a bridging amino acid T
corresponding to amino acid 545 of M78530 PEA-1 P15, a second amino acid sequence being at least 90 % homologous to EKCTVNEECSPSSCLMTEWGEWDECSATCGMGMKKRHRMIKMNPADGSMCKAETSQ
AEKCMMPECHTIPCLLSPWSEWSDCSVTCGKGMRTRQRMLKSLAELGDCNEDLEQVE
KCMLPEC corresponding to amino acids 546 - 665 of Q9HCB6, which also corresponds to amino acids 546 - 665 of M78530 PEA_1 P15, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence RKSWSSSRPITSMFLSPGSPEPASANTARS corresponding to amino acids 666 - 695 of M78530 PEA-1 P15, wherein said first amino acid sequence, bridging amino acid, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M78530 PEA-1 P15, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence RKSWSSSRPITSMFLSPGSPEPASANTARS in M78530 PEA 1 P15.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA_1 P15, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLS corresponding to amino acids 1 - 83 of M78530 PEA_1 P15, a second amino acid sequence being at least 90 % homologous to AAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEEETQFMSNCPVAVTESTPRRRTRIQ
VFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKLCEQDSTFDGVTDKPILDCCACGT
AKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSHSKNYVLWEYGGYASEGVKQVAE
LGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLNVRAAPSAEFSVDRTRHLMSFLTM
MGPSPDWNVGLSAEDLCTKECGW VQKVVQDLIPWDAGTDSGVTYESPNKPTIPQEKIR
PLTSLDHPQSPFYDPEGGSITQVARVVIERIARKGEQCNIVPDNVDDIVADLAPEEKDED
DTPETCIYSNWSPWSACSSSTCDKGKRMRQRMLKAQLDLSVPCPDTQDFQPCMGPGCS
DEDGSTCTMSEWITWSPCSISCGMGMRSRERYVKQFPEDGSVCTLPTEETEKCTVNEEC
SPSSCLMTEWGEWDECSATCGMGMKKRHRM IKMNPADGSMCKAETSQAEKCMMPE
CHTIPCLLSPWSEWSDCSVTCGKGMRTRQRMLKSLAELGDCNEDLEQVEKCMLPEC
corresponding to amino acids 1 - 582 of 094862, which also corresponds to amino acids 84 -665 ofM78530 PEA-1 P15, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence RKSWSSSRPITSMFLSPGSPEPASANTARS corresponding to amino acids 666 - 695 of M78530 PEA-1 P15, wherein said first amino acid sequence, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of M78530 PEA_1 P15, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLS of M78530 PEA 1 P15.
An isolated polypeptide encoding for a tail of M78530 PEA_1 P15, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence RKSWSSSRPITSMFLSPGSPEPASANTARS in M78530 PEA 1 P15.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA-1 P16, comprising a first amino acid sequence being at least 90 % homologous to MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEE
ETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKT.
CEQDSTFDGVTDKPILDCCACGTAKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSH
SKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLN
V corresponding to amino acids 1 - 297 of Q8NCD7, which also corresponds to amino acids 1 -297 of M78530 PEA 1 P16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA-1 P16, comprising a first amino acid sequence being at least 90 % homologous to MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDK VPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEE
ETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTK1CT.
CEQDSTFDGVTDKPILDCCACGTAKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSH
SKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLN
V corresponding to amino acids 1 - 297 of Q9HCB6, which also corresponds to amino acids 1 -297 of M78530 PEA 1 P 16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA_1 P16, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLS corresponding to amino acids 1 - 83 of M78530 PEA 1 P16, and a second amino acid sequence being at least 90 %
homologous to AAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEEETQFMSNCPVAVTESTPRRRTRIQ
VFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKLCEQDSTFDGVTDKPILDCCACGT
AKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSHSKNYVLWEYGGYASEGVKQVAE
LGSPVKMEEEIRQQSDEVLTVIKAKAQWPAWQPLNV corresponding to amino acids 1 -214 of 094862, which also corresponds to amino acids 84 - 297 of M78530 PEA-1 P16, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of M78530 PEA 1 P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLS of M78530 PEA 1 P 16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA_1 P17, comprising a first amino acid sequence being at least 90 % homologous to MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEE
ETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKL
SKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQ corresponding to amino acids I -275 of Q8NCD7, which also corresponds to amino acids I - 275 of M78530 PEA_1 P17, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide 15 having the sequence VRQKNHRMTK corresponding to amino acids 276 - 285 of M78530 PEA-1 P 17, wherein said first amino acid sequence and second amino acid sequence are cont iguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M78530 PEA-1 P17, comprising a polypeptide 20 being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRQKNHRMTK in M78530 PEA 1 P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA-1 P17, comprising a first amino acid 25 sequence being at least 90 % homologous to MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLSAAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEE
ETQFMSNCPVAVTESTPRRRTRIQVFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKT.
CEQDSTFDGVTDKPILDCCACGTAKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSH
30 SKNYVLWEYGGYASEGVKQVAELGSPVKMEEEIRQQ corresponding to amino acids 1 -275 of Q9HCB6, which also corresponds to amino acids 1 - 275 of M78530 PEA_1 P17, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRQKNHRMTK corresponding to amino acids 276 - 285 of M78530 PEA,I P17, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M78530 PEA-1 P17, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRQKNHRMTK in M78530 PEA 1 P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M78530 PEA-1 P17, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYT
EFSLRVEGDPDFYKPGTSYRVTLS corresponding to amino acids 1 - 83 of M78530 PEA_1 P17, a second amino acid sequence being at least 90 % homologous to AAPPSYFRGFTLIALRENREGDKEEDHAGTFQIIDEEETQFMSNCPVAVTESTPRRRTRIQ
VFWIAPPAGTGCVILKASIVQKRIIYFQDEGSLTKKLCEQDSTFDGVTDKPILDCCACGT
AKYRLTFYGNWSEKTHPKDYPRRANHWSAIIGGSHSKNYVLWEYGGYASEGVKQVAE
LGSPVKMEEEIRQQ corresponding to amino acids 1 - 192 of 094862, which also corresponds to amino acids 84 - 275 of M78530 PEA_1 P17, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and rr~st preferably at least 95% homologous to a polypeptide having the sequence VRQKNHRMTK
corresponding to amino acids 276 - 285 of M78530 PEA-1 P17, wherein said first amino acid sequence, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of M78530 PEA-1 P17, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the .seguej~ce MRLSPAPLKLSRTPALLALALPLAAALAFSDETLDKVPKSEGYCSRILRAQGTRREGYTEFSLR
VEGDPDFYKPGTSYRVTLS of M78530 PEA I P17.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M78530 PEA-1 P17, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRQKNHRMTK in M78530 PEA 1 P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T48119 P2, comprising a first amino acid sequence being at least 90 % homologous to MTRQMASSGASGGKIDNSVLVLIVGLSTVGAGAYAYKTMKEDEKRYNERISGLGLTPE
ELPYMRPPLSKELWFSDDPNVTKTLRFKQWNGKERSIYFQPPSFYVSAQDLPHIENGGV
AVLTGKKVVQLDVRDNMVKLNDGSQITYEKCLIATGGTPRSLSAIDRAGAEVKSRTTL
FRKIGDFRSLEKISREVKSITIIGGGFLGSELACALGRKARALGTEVIQLFPEKGNMGKILP
EYLSNWTMEKVRREGVKVMPNAIVQSVGVSSGKLLIKLKDGRKVETDHIVAAVGLEP
NVELAKTGGLEIDSDFGGFRVNAELQARSNIWVAGDAACFYDIKLGRRRVEHHDHAV
VSGRLAGENMTGAAKPYWHQSMFWSDLGPDVGYEAIGLVDSSLPTVGVFAKATAQD
NPKSATEQSGTGIRSESETESEASEITIPPSTPAVPQAPVQGEDYGKGVIFYLRDKVVVGI
VLWNIFNRMPIARKIIKDGEQHEDLNEVAKLFNIHED corresponding to amino acids 50 -613 of PCD8 HI1MAN, which also corresponds to amino acids 1 - 564 of T48119 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T48119 P2, comprising a first amino acid sequence being at least 90 % homologous to MTRQMASSGASGGKIDNSVLVLIVGLSTVGAGAYAYKTMKEDEKRYNERISGLGLTPE
QKQKKAALSASEGEEVPQDKAPSHVPFLLIGGGTAAFAAARSIRARDPGARVLIVSEDP
ELPYMRPPLSKELWFSDDPNVTKTLRFKQWNGKERSIYFQPPSFYVSAQDLPHIENGGV
AVLTGKKVVQLDVRDNMVKLNDGSQITYEKCLIATGGTPRSLSAIDRAGAEVKSRTTL
FRKIGDFRSLEKISREVKSITIIGGGFLGSELACALGRKARALGTEVIQLFPEKGNMGKILP
EYLSNWTMEKVRREGVKVMPNAIVQSVGVSSGKLLIKLKDGRKVETDHIVAAVGLEP
VSGRLAGENMTGAAKPYWHQSMFWSDLGPDVGYEAIGLVDSSLPTVGVFAKATAQD
NPKSATEQSGTGIRSESETESEASEITIPPSTPAVPQAPVQGEDYGKGVIFYLRDKVVVGI
VLWNIFNRMPIARKI.IKDGEQHEDLNEVAKLFNIHED corresponding to amino acids 50 -613 of PCD8~HUMAN, which also corresponds to amino acids 1 - 564 of T48119 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971 P6, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAW VALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
IO KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
LKPEEEAPAPEVGASKPEGIDSRPETLI-IPGRPQPPAEEELCSGKPFDAFTDLKNGSLFAFR
GQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAFTRINCQGKTYLFKGSQYWRFEDGV
LDPDYPRNISDGFDGIPDNVDAALALPAHSYSGRERVYFFKG corresponding to amino acids 1 - 276 of VTNC HUMAN, which also corresponds to amino acids 1 - 276 of 15 T39971 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence TQGVVGD corresponding to amino acids 277 - 283 of T39971 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
20 According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T39971 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TQGWGD
in T39971 P6.
25 According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971 P9, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAWALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
GQYCYELDEKAVRPGYPKLIRDWGIEGPIDAAFTRINCQGKTYLFKGSQYWRFEDGV
LDPDYPRNISDGFDGIPDNVDAALALPAHSYSGRERVYFFKGKQYWEYQFQHQPSQEE
CEGSSLSAVFEI-IFAMMQRDSWEDIFELLFWGRT corresponding to amino acids 1 - 325 of VTNC_HUMAN, which also corresponds to amino acids 1 - 325 of T39971 P9, and a second amino acid sequence being at least 90 % homologous to SGMAPRPSLAKKQRFRI-IRNRKGYRSQRGHSRGRNQNSRRPSRATWLSLFSSEESNLGA
NNYDDYRMDWLVPATCEPIQS VFFFSGDKYYRVNLRTRRV DTV DPPYPRSIAQY W LGC
PAPGHL corresponding to amino acids 357 - 478 of VTNC HUMAN, which also corresponds to amino acids 326 - 447 of T39971 P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T39971 P9, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise TS, having a structure as follows: a sequence starting from any of amino acid numbers 325-x to 325; and ending at any of amino acid numbers 326 + ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971_PI 1, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAWVALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
LKPEEEAPAPEVGASKPEGIDSRPETLHPGRPQPPAEEELCSGKPFDAFTDLKNGSLFAFR
GQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAFTRINCQGKTYLFKGSQYWRFEDGV
LDPDYPRNISDGFDGIPDNVDAALALPAHSYSGRERVYFFKGKQYWEYQFQHQPSQEE
CEGSSLSAVFEHFAMMQRDSWEDIFELLFWGRTS corresponding to amino acids 1 - 326 of VTNC_HUMAN, which also corresponds to amino acids 1 - 326 ofT39971 P11, and a second amino acid sequence being at least 90 % homologous to DKYYRVNLRTRRVDTVDPPYPRSIAQYWLGCPAPGHL corresponding to amino acids 442 - 478 of VTNC_HUMAN, which also corresponds to amino acids 327 - 363 of T39971 P1 I, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T39971 P 11, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more 5 preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise SD, having a structure as follows: a sequence starting from any of amino acid numbers 326-x to 326; and ending at any of amino acid numbers 327 + ((rr2) - x), in which x varies from 0 to rr2.
10 According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971 Pl I, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAWVALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
I S LKPEEEAPAPEVGASKPEGIDSRPETLHPGRPQPPAEEELCSGKPFDAFTDLKNGSLFAFR
GQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAFTRINCQGKTYLFKGSQYWRFEDGV
LDPDYPRNISDGFDGIPDNVDAALALPAHSYSGRERVYFFKGKQYWEYQFQHQPSQEE
CEGSSLSAVFEHFAMMQRDSWEDIFELLFWGRTS corresponding to amino acids 1 - 326 of Q9BSH7, which also corresponds to amino acids 1 - 326 of T39971 PI 1, and a second amino 20 acid sequence being at least 90 % homologous to DKYYRVNLRTRRVDTVDPPYPRSIAQYWLGCPAPGHL corresponding to amino acids 442 - 478 of Q9BSH7, which also corresponds to amino acids 327 - 363 of T39971 P11, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an 25 isolated chimeric polypeptide encoding for an edge portion of T39971 P11, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino . acids in length, wherein at least two amino acids comprise SD, having a structure as follows: a 30 sequence starting from any of amino acid numbers 326-x to 326; and ending at any of amino acid numbers 327 + ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971 P12, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAWVALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
LKPEEEAPAPEVGASKPEGIDSRPETLHPGRPQPPAEEELCSGKPFDAFTDLKNGSLFAFR
GQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAFTRINCQGKTYLFK corresponding to amino acids 1 - 223 of VTNC HUMAN, which also corresponds to amino acids 1 -223 of T39971 P12, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence VPGAVGQGRKHLGRV corresponding to amino acids 224 - 238 of T39971 P12, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T39971 P12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VPGAVGQGRKHLGRV in T39971 P12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T39971 P12, comprising a first amino acid sequence being at least 90 % homologous to MAPLRPLLILALLAWVALADQESCKGRCTEGFNVDKKCQCDELCSYYQSCCTDYTAEC
KPQVTRGDVFTMPEDEYTVYDDGEEKNNATVHEQVGGPSLTSDLQAQSKGNPEQTPV
LKPEEEAPAPEVGASKPEGIDSRPETLHPGRPQPPAEEELCSGKPFDAFTDLKNGSLFAFR
GQYCYELDEKAVRPGYPKLIRDVWGIEGPIDAAFTRINCQGKTYLFK corresponding to amino acids 1 - 223 of Q9BSH7, which also corresponds to amino acids 1 - 223 of T39971 P12, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VPGAVGQGRKHLGRV corresponding to amino acids 238 of T39971 P12, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T39971 P12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VPGAVGQGRKHLGRV in T39971 P12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 244808 PEA-1 P5, comprising a first amino acid -sequence being at least 90 % homologous to MLLPQLCWLPLLAGLLPPV.PAQKFSALTFLRVDQDKDKDCSLDCAGSPQKPLCASDGR
TFLSRCEFQRAKCKDPQLEIAYRGNCKDVSRCVAERKYTQEQARKEFQQVFIPECNDD
GTYSQVQCHSYTGYCWCVTPNGRPISGTAVAHKTPRCPGSVNEKLPQREGTGKTDDAA
APALETQPQGDEEDIASRYPTLWTEQVKSRQNKTNKNSVSSCDQEHQSALEEAKQPKN
DNWIPECAHGGLYKPVQCHPSTGYCWCVLVDTGRPIPGTSTRYEQPKCDNTARAHPA
KARDLYKGRQLQGCPGAKKHEFLTSVLDALSTDMVHAASDPSSSSGRLSEPDPSHTLEE
RVVHWYFKLLDKNSSGDIGKKEIKPFKRFLRKKSKPKKCVKKFVEYCDVNNDKSISVQ
ELMGCLGVAKEDGKADTKKRHTPRGHAESTSNRQ corresponding to amino acids 1 - 441 of SM02 HUMAN, which also corresponds to amino acids 1 - 441 of 244808 PEA-1 PS, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DAMVVSSRPKATTHRKSRTLSRR corresponding to amino acids 442 - 464 of 244808 PEA-1 P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 244808 PEA_1 P5, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DAMVVSSRPKATTHRKSRTLSRR in 244808 PEA 1 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 244808 PEA-1 P6, comprising a first amino acid sequence being at least 90 % homologous to MLLPQLCWLPLLAGLLPPVPAQKFSALTFLRVDQDKDKDCSLDCAGSPQKPLCASDGR
TFLSRCEFQRAKCKDPQLEIAYRGNCKDVSRCVAERKYTQEQARKEFQQVFIPECNDD
GTYSQVQCHSYTGYCWCVTPNGRPISGTAVAHKTPRCPGS VNEKLPQREGTGKTDDAA
APALETQPQGDEEDIASRYPTLWTEQVKSRQNKTNKNSVSSCDQEHQSALEEAKQPKN
DNVVIPECAHGGLYKPVQCI-IPSTGYCWCVLVDTGRPIPGTSTRYEQPKCDNTARAHPA
KARDLYKGRQLQGCPGAKKHEFLTSVLDALSTDMVHAASDPSSSSGRLSEPDPSHTLEE
RV VH WYFKLLDKNSSGDIGKKEIKPFKRFLRKKSKPKKCVKKFV EYCD VNNDKSISVQ
ELMGCLGVAKEDGKADTKKRH corresponding to amino acids 1 - 428 of SM02_HUMAN, which also corresponds to amino acids 1 - 428 of Z44808,PEA 1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence RSKRNL corresponding to amino acids 429 - 434 of 244808 PEA-1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 244808 PEA_I P6, comprising a polypeptide being I S at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence RSKRNL
in 244808 PEA I P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 244808 PEA-1 P7, comprising a first amino acid sequence being at least 90 % homologous to MLLPQLCWLPLLAGLLPPVPAQKFSALTFLRVDQDKDKDCSLDCAGSPQKPLCASDGR
TFLSRCEFQRAKCKDPQLEIAYRGNCKDVSRCVAERKYTQEQARKEFQQVFIPECNDD
GTYSQVQCHSYTGYCWCVTPNGRPISGTAVAHKTPRCPGSVNEKLPQREGTGKTDDAA
APALETQPQGDEEDIASRYPTLWTEQVKSRQNKTNKNSVSSCDQEHQSALEEAKQPKN
DNVVIPECAHGGLYKPVQCHPSTGYCWCVLVDTGRPIPGTSTRYEQPKCDNTARAHPA
KARDLYKGRQLQGCPGAKKHEFLTSVLDALSTDMVHAASDPSSSSGRLSEPDPSHTLEE
RVVHWYFKLLDKNSSGDIGKKEIKPFKRFLRKKSKPKKCVKKFVEYCDVNNDKSISVQ
ELMGCLGVAKEDGKADTKKRHTPRGHAESTSNRQ corresponding to amino acids 1 - 441 of SM02 HUMAN, which also corresponds to amino acids 1 - 441 of 244808 PEA 1 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a '7 9 polypeptide having the sequence LLWLRGKVSFYCF corresponding to amino acids 442 of 244808 PEA-1 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 244808 PEA_1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence LLWLRGKVSFYCF in 244808 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 244808 PEA_1 P11, comprising a first amino acid sequence being at least 90 % homologous to MLLPQLCWLPLLAGLLPPVPAQKFSALTFLRVDQDKDKDCSLDCAGSPQKPLCASDGR
TFLSRCEFQRAKCKDPQLEIAYRGNCKDVSRCVAERKYTQEQARKEFQQVFIPECNDD
GTYSQVQCHSYTGYCWCVTPNGRPISGTAVAHKTPRCPGSVNEKLPQREGTGKT
1 S corresponding to amino acids 1 - 170 of SM02 HUMAN, which also corresponds to amino acids 1 - 170 of 244808 PEA_1 P11, and a second amino acid sequence being at least 90 homologous to DIASRYPTLWTEQVKSRQNKTNKNSVSSCDQEHQSALEEAKQPKNDNV VIPECAHGGL
YKPVQCHPSTGYCWCVLVDTGRPIPGTSTRYEQPKCDNTARAHPAKARDLYKGRQLQ
GCPGAKKHEFLTSVLDALSTDMVHAASDPSSSSGRLSEPDPSHTLEERVVHWYFKLLD
KNSSGDIGKKEIKPFKRFLRKKSKPKKCVKKFVEYCDVNNDKSISVQELMGCLGVAKE
DGKADTKKRHTPRGHAESTSNRQPRKQG corresponding to amino acids 188 - 446 of SM02 HUMAN, which also corresponds to amino acids 171 - 429 of 244808 PEA,1 P1 l, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of 244808 PEA-1 P11, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise TD, having a structure as follows: a sequence starting from any of amino acid numbers 170-x to -170; and ending at any of amino acid numbers 171+ ((,r2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for S67314 PEA_I P4, comprising a first amino acid 5 sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence M VDAFLGTWKLV DSKNFDDYMKSLGVGFATRQVASMTKPTTIIEKNGDILTLKTHSTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of FABH HUMAN, which also corresponds to amino 10 acids 1 - 116 of 567314 PEA-1 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence VRWATLELYLIGYYYCSFSQACSKKPSPPLRAVEAGTREWLWVRVVSGGNFLCSGFGL
TQAGTQILPYRLHDCGQITFSKCNCKTGINNTNLVGLLGSL corresponding to amino acids 15 117 - 215 of 567314 PEA_1 P4, wherein said firstand second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA-1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at 20 least about 90% and most preferably at least about 95% homologous to the sequence VRWATLELYLIGYYYCSFSQACSKKPSPPLRAVEAGTREWLWVRVVSGGNFLCSGFGL
TQAGTQILPYRLHDCGQITFSKCNCKTGINNTNLVGLLGSL in 567314 PEA 1 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 567314 PEA_1 P4, comprising a first amino acid 25 sequence being at least 90 % homologous to MVDAFLGTWKLVDSKNFDDYMKSLGVGFATRQVASMTKPTTIIEKNGDILTLKTHSTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of AAP35373, which also corresponds to amino acids 1 -116 of 567314 PEA-1 P4, and a second amino acid sequence being at least 70%, optionally at 30 least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRWATLELYLIGYYYCSFSQACSKKPSPPLRAVEAGTREWLWVRVVSGGNFLCSGFGL
TQAGTQIL.PYRLHDCGQITFSKCNCKTGINNTNLVGLLGSL corresponding to amino acids 117 - 215 of 567314 PEA_1 P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA_1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRWATLELYLIGYYYCSFSQACSKKPSPPLRAVEAGTREWLWVRVVSGGNFLCSGFGL
TQAGTQILPYRLHDCGQITFSKCNCKTGINNTNLVGLLGSL in 567314 PEA_I P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 567314 PEA_1 P5, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MVDAFLGTWKLVDSKNFDDYMKSLGVGFATRQVASMTKPTTIIEKNGDILTLKTHSTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of FABH HUMAN, which also corresponds to amino acids 1 - 116 of S67314 PEA_1 P5, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DVLTAWPSIYRRQVKVLREDEITILPWHLQWSREKATKLLRPTLPSYNNHGWEELRVG
KSIV corresponding to amino acids 117 - 178 of S67314 PEA_1 P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA_1 P5, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DVLTAWPSIYRRQVKVLREDEITILPWHLQWSREKATKLLRPTLPSYNNHGWEELRVG
KSN in 567314 PEA 1 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 567314 PEA-1 P5, comprising a first amino acid' sequence being at least 90 % homologous to MVDAFLGTWKLVDSKNFDDYMKSLGVGFATRQVASMTKPTTI f EKNGDILTLKTI-ISTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of AAP35373, which also corresponds to amino acids 1 116 of 567314 PEA-1 P5, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DVLTAWPSIYRRQVKV LREDEITILPWHLQWSREKATKLLRPTLPSYNNHGWEELRVG
KSIV corresponding to amino acids 117 - 178 of S67314 PEA-1 PS, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA-1 PS, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DVLTAWPSIYRRQVKVLREDEITILPWHLQWSREKATKLLRPTLPSYNNHGWEELRVG
KSIV in S67314 PEA 1 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimerie polypeptide encoding for S67314 PEA-1 P6, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MVDAFLGTWKLVDSKNFDDYMKSLGVGFATRQVASMTKPTTIIEKNGDILTLKTHSTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of FABH HUMAN, which also corresponds to amino acids 1 - 116 of S67314 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence MEKLQLRNVK
corresponding to amino acids 117 - 126 of 567314 PEA_1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MEKLQLRNVK in S67314 PEA-1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 567314 PEA-1 P6, comprising a first amino acid sequence being at least 90 % homologous to MVDAFLGTWKLVDSKNFDDYM.KSLGVGFATRQVASMTKPTTIIEKNGDILTLKTHSTF
KNTEISFKLGVEFDETTADDRKVKSIVTLDGGKLVHLQKWDGQETTLVRELIDGKLIL
corresponding to amino acids 1 - 116 of AAP35373, which also corresponds to amino acids 1 -116 of S67314 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MEKLQLRNVK corresponding to amino acids 117 - 126 of 567314 PEA-1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 567314 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MEKLQLRNVK in S67314 PEA 1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 567314 PEA-1 P7, comprising a first amino acid sequence being at least 90 % homologous to MVDAFLGTWKLVDSKNFDDYMKSL
corresponding to amino acids 1 - 24 of FABH HUMAN, which also corresponds to amino acids 1 - 24 of 567314 PEA_l P7, second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence AHILITFPLPS corresponding to amino acids 25 - 35 of 567314 PEA 1 P7, and a third amino acid sequence being at least 90 homologous to GVGFATRQVASMTKPTTIIEKNGDILTLKTHSTFKNTEISFKLGVEFDETTADDRKVKSI
VTLDGGKLVHLQKWDGQETTLVRELIDGKLILTLTHGTAVCTRTYEKEA corresponding to amino acids 25 - 133 of FABH HUMAN, which also corresponds to amino acids 36 - 144 of 567314 PEA-1 P7, wherein said first, second, third and fourth amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of S67314 PEA_1 P7, comprising an amino acid sequence being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence encoding for AHILITFPLPS, corresponding to S67314 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for S67314 PEA-1 P7, corr~rising a first amino acid sequence being at least 90 % homologous to MVDAFLGTWKLVDSKNFDDYMKSL
corresponding to amino acids 1 - 24 of AAP35373, which also corresponds to amino acids 1 -24 of S67314 PEA-1 P7, second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence AHILITFPLPS corresponding to amino acids 25 - 35 of 567314 PEA-I P7, and a third amino acid sequence being at least 90 % homologous to GVGFATRQVASMTKPTTIIEKNGDILTLKTHSTFKNTEISFKLGVEFDETTADDRKVKSI
VTLDGGKLVHLQKWDGQETTLVRELIDGKLILTLTHGTAVCTRTYEKEA corresponding to amino acids 25 - 133 of AAP35373, which also corresponds to amino acids 36 -144 of 567314 PEA-1 P7, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of S67314 PEA 1 P7, comprising an amino acid sequence being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence encoding for AHILITFPLPS, corresponding to 567314 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 239337 PEA 2 PEA-1 P4, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWLPLSGAA corresponding to amino acids 1 - 9 of 239337 PEA 2 PEA-1 P4, and a second amino acid sequence being at least 90 homologous to MKKLMVVLSLIAAAWAEEQNKLVHGGPCDKTSHPYQAALYTSGHLLCGGVLIHPLWV
LTAAHCKKPNLQVFLGKHNLRQRESSQEQSSVVRAVIHPDYDAASHDQDIMLLRLARP
S AKLSELIQPLPLERDCSANTTSCHILGWGKTADGDFPDTIQCAYIHLVSREECEHAYPGQ
YTNWIQKTIQAK corresponding to amino acids 1 - 244 of KLK6 HUMAN, which also corresponds to amino acids 10 - 2S3 of 239337 PEA 2 PEA_1 P4, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
10 According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of 239337 PEA 2 PEA-1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 8S%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence MWLPLSGAA of 239337 PEA 2 PEA-1 P4.
1 S According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 239337 PEA 2 PEA-1 P9, comprising a first amino acid sequence being at least 90 % homologous to MKKLMVVLSLIAAAWAEEQNKLVHGGPCDKTSHPYQAALYTSGHLLCGGVLIHPLWV
LTAAHCKKPNLQVFLGKHNLRQRESSQEQSSVVRAVIHPDYDAASHDQDIMLLRLARP
20 AKLSELIQPLPLERDCSANTTSCHILGWGKTADG corresponding to amino acids 1 - 149 of KLK6_HUMAN, which also corresponds to amino acids 1 - 149 of 239337 PEA 2 PEA-1 P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 8S%, more preferably at least 90% and most preferably at least 9S% homologous to a polypeptide having the sequence Q corresponding to amino acids 1 SO -2S 1 SO of 239337 PEA 2 PEA-1 P9, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P10, comprising a first amino acid sequence being at least 90 % homologous to FYYNISE corresponding to amino acids 1 - 67 of PLTP HUMAN, which also corresponds to amino acids I - 67 of HUMPHOSLIP PEA 2 PIO, and a second amino acid sequence being at least 90 % homologous to KVYDFLSTFITSGMRFLLNQQICPVLYHAGTVLLNSLLDTVPVRS SVDELVGIDYSLMK
DPVASTSNLDMDFRGAFFPLTERNWSLPNRAVEPQLQEEERMVYVAFSEFFFDSAMES
YFRAGALQLLLVGDKVPHDLDMLLRATYFGSIVLLSPAVIDSPLKLELRVLAPPRCTIKP
SGTTISVTASVTIALVPPDQPEVQLSSMTMDARLSAKMALRGKALRTQLDLRRFRIYSN
HSALESLALIPLQAPLKTMLQIGVMPMLNERTWRGVQIPLPEGINFVHEVVTNHAGFLTI
GADLHFAKGLREVIEKNRPADVRASTAPTPSTAAV corresponding to amino acids 163 -493 of PLTP HUMAN, which also corresponds to amino acids 68 - 398 of HUMPHOSLIP PEA 2 P10, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMPHOSLIP PEA 2 P10, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EK, having a structure as follows: a sequence starting from any of amino acid numbers 67-x to 67; and ending at any of amino acid numbers 68+ ((n-2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P12, comprising a first amino acid sequence being at least 90 % homologous to MALFGALFLALLAGAHAEFPGCKIRVTSKALELVKQEGLRFLEQELETITIPDLRGKEGH
FYYNISEVKVTELQLTSSELDFQPQQELMLQITNASLGLRFRRQLLYWFFYDGGYINAS
AEGVSIRTGLELSRDPAGRMKVSNVSCQASVSRMHAAFGGTFKKVYDFLSTFITSGMRF
LLNQQICPVLYHAGTVLLNSLLDTVPVRSSVDELVGIDYSLMKDPVASTSNLDMDFRG
AFFPLTERNWSLPNRAVEPQLQEEERMVYVAFSEFFFDSAMESYFRAGALQLLLVGDK
VPHDLDMLLRATYFGSIVLLSPAVIDSPLKLELRVLAPPRCTIKPSGTTISVTASVTIALVP
PDQPEVQLSSMTMDARLSAKMALRGKALRTQLDLRRFRIYSNHSALESLALIPLQAPLK
TMLQIGVMPMLN corresponding to amino acids 1 - 427 of PLTP HUMAN, which also corresponds to amino acids 1 - 427 of HUMPHOSLIP PEA 2 P12, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85'%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence i, GKAGV corresponding to amino acids 428 - 432 of HUMPI-IOSLIP PEA 2 P 12, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMPHOSLIP PEA,2 P 12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence GKAGV in HUMPHOSLIP~PEA 2 P 12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P31, comprising a first amino acid sequence being at least 90 % homologous to MALFGALFLALLAGAHAEFPGCKIRVTSKALELVKQEGLRFLEQELETITIPDLRGKEGH
I 5 FYYNISE corresponding to amino acids 1 - 67 of PLTP HUMAN, which also corresponds to amino acids 1 - 67 of HUMPHOSLIP PEA 2 P31, and a second amino acid sequence being at .yp: _ least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PGLERGADKFPVVGGSSLFLALDLTLRPPVG corresponding to amino acids 68 - 98 of HUMPHOSLIP PEA 2 P31, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMPHOSLIP PEA 2 P31, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence PGLERGADKFPVVGGSSLFLALDLTLRPPVG in HUMPHOSLIP PEA 2 P31.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P33, comprising a first amino acid sequence being at least 90 % homologous to MALFGALFLALLAGAHAEFPGCKIRVTSKALELVKQEGLRFLEQELETITIPDLRGKEGH
FYYNISEVKVTELQLTSSELDFQPQQELMLQITNAS LGLRFRRQLLYWFFYDGGYINAS
AEGVSIRTGLELSRDPAGRMKVSNVSCQASVSRMHAAFGGTFKKVYDFLSTFITSGMRF
LLNQQ corresponding to amino acids 1 - 183 of PLTP HUMAN, which also corresponds to amino acids 1 - 183 of HUMPHOSLIP PEA 2 P33, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VWAATGRRVARVGMLSL corresponding to amino acids 184 - 200 of HUMPHOSLIP PEA 2 P33, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMPHOSLIP PEA 2 P33, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence VWAATGRRVARVGMLSL in HUMPHOSLIP PEA 2 P33.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P34, comprising a first amino acid sequence being at least 90 % homologous to MALFGALFLALLAGAHAEFPGCKIRVTSKALELVKQEGLRFLEQELETITIPDLRGKEGH
FYYNISEVKVTELQLTSSELDFQPQQELMLQITNASLGLRFRRQLLYWFFYDGGYINAS
AEGVSIRTGLELSRDPAGRMKVSNVSCQASVSRMHAAFGGTFKKVYDFLSTFITSGMRF
LLNQQICPVLYHAGTVLLNSLLDTVPV corresponding to amino acids 1 - 205 of PLTP HUMAN, which also corresponds to amino acids 1 - 205 of HUMPHOSLIP PEA 2 P34, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence LWTSLLALTIPS corresponding to amino acids 206 - 217 of HUMPHOSLIP PEA 2 P34, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMPHOSLIP PEA 2 P34, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence LWTSLLALTIPS in HUMPHOSLIP PEA 2 P34.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMPHOSLIP PEA 2 P35, comprising a first amino acid sequence being at least 90 % homologous to MALFGALFLALLAGAI IAEFPGCKIRVTSKALELVKQEGLRFLEQELETITIPDLRGKEGH
FYYNISEVKVTELQLTSSELDFQPQQELMLQITNASLGLRFRRQLLYWF corresponding to amino acids I - 109 of PLTP HUMAN, which also corresponds to amino acids 1 -109 of HUMPHOSLIP PEA 2 P35, a second amino acid sequence bridging amino acid sequence comprising of L, a third amino acid sequence being at least 90 % homologous to KVYDFLSTFITSGMRFLLNQQ corresponding to amino acids I 63 - 183 of PLTP HUMAN, which also corresponds to amino acids 111 - 131 of HUMPHOSLIP PEA 2 P35, and a fourth amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VWAATGRRVARVGMLSL corresponding to amino acids 132 - 148 of HUMPHOSLIP PEA 2 P35, wherein said first amino acid sequence, second amino acid sequence, third amino acid sequence and fourth amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of HUMPHOSLIP PEA 2 P35, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise FLK
having a structure as follows (numbering according to HUMPHOSLIP PEA 2 P35): a sequence starting from any of amino acid numbers 109-x to 109; and ending at any of amino acid numbers 111 +
((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMPHOSLIP PEA 2 P35, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence VWAATGRRVARVGMLSL in HUMPHOSLIP PEA 2 P35.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832-P7, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNG corresponding to amino acids 12 - 223 of GILT HUMAN, which also corresponds to amino acids 1 - 212 of T59832 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, 10 more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRIFLALSLTLIVPWSQGWTRQRDQR corresponding to amino acids 213 - 238 of T59832 P7, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an 15 isolated polypeptide encoding for a tail of T59832 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRIFLALSLTLIVPWSQGWTRQRDQR in T59832 P7.
According to preferred embodiments of the present invention, there is provided an 20 isolated chimeric polypeptide encoding for T59832 P9, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
25 ECAMGDRGMQLMHANAQRTDALQPPHE corresponding to amino acids 12 - 214 of GILT HUMAN, which also corresponds to amino acids 1 - 203 of T59832 P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR corresponding to 30 amino acids 204 - 244 of T59832 P9, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80°I°, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR in T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P12, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVE corresponding to amino acids 12 - 141 of GILT HUMAN, which also corresponds to amino acids 1 - 130 of T59832 P12, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGD RGMQLMHANAQRTDALQPPHEYVPW VTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 173 - 261 of GILT HUMAN, which also corresponds to amino acids 131 - 219 of T59832 P12, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P12, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EC, having a structure as follows: a sequence starting from any of amino acid numbers 130-x to 130; and ending at any of amino acid numbers 131+ ((n-2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P18, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYK corresponding to amino acids 12 - 55 of GILT HUMAN, which also corresponds to amino acids I - 44 of T59832 P18, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 173 - 261 of GILT HUMAN, which also corresponds to amino acids 45 - 133 of T59832 P 18, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P18, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise KC, having a structure as follows: a sequence starting from any of amino acid numbers 44-x to 44; and ending at any of amino acid numbers 45+ ((rr2) - x), in which x varies from 0 to rr2.
I 5. According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P4, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVWHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWLFGMGNEVDVH
AAFFHGQALTNKNYRIDTINLFPATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFF
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTWSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSWWL
FSAGNEADVHGIYFSGNTYLWRGERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDH
YTGGMKQKYTVNQCRRQSEDSTFYLGERTWIAAVEVEWDYSPQREWEKELHHLQEQ
NVSNAFLDKGEFYIGSKYKKVVYRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDK
YSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYS
DHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHT
V H FHGHSFQYKHRGVYSSDVFDIFPGTYQTLEMFPRTPGIWLLHCHVTDHIHAGMETT
YTVLQNE corresponding to amino acids 1 - 1060 of CERU HUMAN, which also corresponds to amino acids 1 - 1060 of HSCP2 PEA-1 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GGTSM
corresponding to amino acids 1061 - 1065 of HSCP2 PEA_1 P4, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA_1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GGTSM in HSCP2 PEA 1 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P8, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKL,ISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
AAFFHGQALTNKNYRIDTINLFPATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFF
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSVV WYL
FSAGNEADVHGIYFSGNTYLWRGERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDH
YTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQ
NVSNAFLDKGEFYIGSKYKKVVYRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDK
VKIIFKNMATRPYSIHAHGVQTESSTVTPTLPGETLTYVWKIPERSGAGTEDSACIPWAY
YSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYS
DHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHT
VHFHGHSFQYK corresponding to amino acids 1 - 1006 ofCERU HUMAN, which also corresponds to amino acids 1 - 1006 of HSCP2 PEA-1 P8, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence KCFQEHLEFGYSTAM corresponding to amino acids 1007 - 1021 of HSCP2 PEA-1 P8, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA_1 P8, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KCFQEHLEFGYSTAM in HSCP2 PEA-1 P8.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA_1 P14, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
AAFFHGQALTNKNYRIDTINLFPATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFF
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMH corresponding to amino acids 1 - 621 of CERU HUMAN, which also corresponds to amino acids 1 - 621 of HSCP2_PEA-1 P14, a second amino acid sequence bridging amino acid sequence comprising of W, and a third amino acid sequence being at least 90 % homologous to TFNVECLTTDHYTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQRE
WEKELHHLQEQNVSNAFLDKGEFYIGSKYKKVVYRQYTDSTFRVPVERKAEEEHLGIL
GTEDSACIPWAYYSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENE
SWYLDDNIKTYSDHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMH V GDEVN WYL
MGMGNEIDLHTVHFHGHSFQYKHRGVYSSDVFDIFPGTYQTLEMFPRTPGI W LLHCHV
TDHIHAGMETTYTVLQNEDTKSG corresponding to amino acids 694 - 1065 of 10 CERU HUMAN, which also corresponds to amino acids 623 - 994 of HSCP2 PEA-1 P14, wherein said first amino acid sequence, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of HSCP2 PEA_1 P14, comprising a 15 polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise HWT having a structure as follows (numbering according to HSCP2 PEA_1 P14): a sequence starting from any of amino acid 20 numbers 621-x to 621; and ending at any of amino acid numbers 623 + ((rr2) -x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA 1 P15, comprising a first amino acid sequence being at least 90 % homologous to RIGRLYKKALYLQYTDETFRTTIEKPV WLGFLGPIIKAETGDKVWHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFWMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSVVWYL
FSAGNEADVHGIYFSGNTYLWRGERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDH
YTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQ
NV SNAFLDKGEFYIGSKYKKWYRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDK
VKIIFKNMATRPYSIHAHGVQTESSTVTPTLPGETLTYVWKIPERSGAGTEDSACIPWAY
YSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYS
DHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHT
VHFHGHSFQYKHRGVYSSDVFDIFPGTYQTLEMFPRTPGIWLLHCI-IVTDHIHAGMETT
YTVLQNE corresponding to amino acids I - 1060 of CERU I-IUMAN, which also corresponds to amino acids 1 - 1060 of HSCP2 PEA-1 P15, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and I S most preferably at least 95% homologous to a polypeptide having the sequence GEYPASSETHRRIWNVIYPITVSVIILFQISTKE corresponding to amino acids 1061 - 1094 of HSCP2 PEA_1 P15, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA-1 P 15, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GEYPASSETHRRIWNVIYPITVSVIILFQISTKE in HSCP2 PEA_1 P15.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P2, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFWMFSWDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
AAFFHGQALTNKNYRIDTINLFPATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFF
QV QECN:KSSSKDNIRGKHV RHYYIAAEEIIWNYAPSGIDIFTKEN LTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSI EPIG VRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNI RMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSVVWYL
YTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQ
corresponding to amino acids 1 - 761 of CERU HUMAN, which also corresponds to amino acids 1 - 761 of HSCP2 PEA_1 P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence K
corresponding to amino acids 762 - 762 of HSCP2 PEA_1 P2, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA,1 P16, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSWDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
AAFFHGQALTNKNYRIDTINLFPATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFF
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLWREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSWWYL
FSAGNEADVHGIYFSGNTYLWRGERRDTAN LFPQTSLTLHMWDTEGTFNVECLTTDH
YTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQ
NVSNAFLDKGEFYIGSKYKKVWRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDK
V KI IFKNMATRPYSIHAHG VQTESSTVTPTLPGETLTYV W KIPERSGAGTEDSACIPWA Y
YSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYS
DHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHT
VHFHGHSFQYKH corresponding to amino acids 1 - 1007 of CERU HUMAN, which also corresponds to amino acids 1 - 1007 of HSCP2 PEA_1 P16, and a second amino acid sequence being at least 70%, optiona lly at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence LLRLTGEYGM corresponding to amino acids 1008 - 1017 of HSCP2 PEA_1 P16, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA_I P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence IS LLRLTGEYGM in HSCP2 PEA 1 P16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P6, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMHSMNGFMYGNQPGLTMCKGDSVVWYL
FSAGNEADVHGIYFSGNTYLWRGERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDH
YTGGMKQKYTVNQCRRQSEDSTFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQ
VKIIFKNMATRPYSIHAHGVQTESSTVTPTLPGETLTYVWKIPERSGAGTEDSACIPWAY
YSTVDQVKDLYSGLIGPLIVCRRPYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYS
DHPEKVNKDDEEFIESNKMHAINGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHT
VHFHGHSFQYK corresponding to amino acids 1 - 1006 of CERU HUMAN, which also corresponds to amino acids 1 - 1006 of HSCP2 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GSL
corresponding to amino acids 1007 - 1009 of HSCP2 PEA_1 P6, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA_l P22, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHE corresponding to amino acids 1 - 131 of CERU HUMAN, which also corresponds to amino acids 1 - 131 of HSCP2 PEA-1 P22, a second amino acid sequence bridging amino acid sequence comprising of A, and a third amino acid sequence~being at least 90 % homologous to ATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFFQVQECNKSSSKDNIRGKHVRHY
YIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQGTTRIGGSYKKLVYREYTDASFTN
RKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAYPLSIEPIGVRFNKNNEGTYYSPNY
NPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNADPVCLAKMYYSAVDPTKDIFTGLI
GPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENESLLLEDNIRMFTTAPDQVDKEDE
DFQESNKMHSMNGFMYGNQPGLTMCKGDSWWYLFSAGNEADVHGIYFSGNTYLWR
GERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDHYTGGMKQKYTVNQCRRQSEDS
TFYLGERTYYIAAVEVEWDYSPQREWEKELHHLQEQNVSNAFLDKGEFYIGSKYKKW
YRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDKVKIIFKNMATRPYSIHAHGVQTE
SSTVTPTLPGETLTYVWKIPERSGAGTEDSACIPWAYYSTVDQVKDLYSGLIGPLIVCRR
PYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYSDHPEKVNKDDEEFIESNKMHAI
NGRMFGNLQGLTMI-IVGDEVNWYLMGMGNEIDLHTVHFI-IGHSFQYKHRGVYSSDVF
DIFPGTYQTLEMFPRTPGIWLLHCHVTDHIHAGMETTYTVLQNEDTKSG corresponding to amino acids 262 - 1065 of CERU HUMAN, which also corresponds to amino acids 936 of HSCP2 PEA_1 P22, wherein said first amino acid sequence, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of HSCP2_PEA-1 P22, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EAV having a structure as follows (numbering according to HSCP2 PEA-1 P22): a sequence starting from any of amino acid numbers 131-x to 131; and ending at any of amino acid numbers 133 + ((n-2) -x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA_I P24, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MPLTMGKRNLFLLTP corresponding to amino acids 1 - 15 of HSCP2 PEA_1 P24, and a second amino acid sequence being at least 90 % homologous to VNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVHAAFFHGQALTNKNYRIDTINLFP
ATLFDAYMVAQNPGEWMLSCQNLNHLKAGLQAFFQVQECNKSSSKDNIRGKHVRHY
YIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQGTTRIGGSYKKLWREYTDASFTN
RKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAYPLSIEPIGVRFNKNNEGTWSPNY
NPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNADPVCLAKMYYSAVDPTKDIFTGLI
GPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENESLLLEDNIRMFTTAPDQVDKEDE
DFQESNKMHSMNGFMYGNQPGLTMCKGDSVVWYLFSAGNEADVHGIYFSGNTYLWR
GERRDTANLFPQTSLTLHMWPDTEGTFNVECLTTDHYTGGMKQKYTVNQCRRQSEDS
TFYLGERTWIAAVEVEWDYSPQREWEKELHHLQEQNVSNAFLDKGEFYIGSKYKKW
YRQYTDSTFRVPVERKAEEEHLGILGPQLHADVGDKVKIIFKNMATRPYSIHAHGVQTE
SSTVTPTLPGETLTYV WKIPERSGAGTEDSACIPWAYYSTVDQVKDLYSGLIGPLIVCRR
PYLKVFNPRRKLEFALLFLVFDENESWYLDDNIKTYSDHPEKVNKDDEEFIESNKMHAI
NGRMFGNLQGLTMHVGDEVNWYLMGMGNEIDLHTVHFI-IGHSFQYKHRGVYSSDVF
DIFPGTYQTLEMFPRTPGIWLLHCHVTDHIHAGMETTYTVLQNEDTKSG corresponding to amino acids 262 - 1065 of CERU HUMAN, which also corresponds to amino acids of HSCP2 PEA-1 P24, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of HSCP2 PEA-1 P24, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MPLTMGKRNLFLLTP of HSCP2 PEA 1 P24.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P25, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPV WLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HGITYYKEHEGAIYPDNTTDFQRADDKVYPGEQYTYMLLATEEQSPGEGDGNCVTRIY
HSHIDAPKDIASGLIGPLIICKKDSLDKEKEKHIDREFVVMFSVVDENFSWYLEDNIKTY
CSEPEKVDKDNEDFQESNRMYSVNGYTFGSLPGLSMCAEDRVKWYLFGMGNEVDVH
QVQECNKSSSKDNIRGKHVRHYYIAAEEIIWNYAPSGIDIFTKENLTAPGSDSAVFFEQG
TTRIGGSYKKLVYREYTDASFTNRKERGPEEEHLGILGPVIWAEVGDTIRVTFHNKGAY
PLSIEPIGVRFNKNNEGTYYSPNYNPQSRSVPPSASHVAPTETFTYEWTVPKEVGPTNAD
PVCLAKMYYSAVDPTKDIFTGLIGPMKICKKGSLHANGRQKDVDKEFYLFPTVFDENES
LLLEDNIRMFTTAPDQVDKEDEDFQESNKMH corresponding to amino acids 1 - 621 of CERU HUMAN, which also corresponds to amino acids 1 - 621 of HSCP2 PEA-1 P25, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence CKYCIIHQSTKLF corresponding to amino acids 622 - 634 of HSCP2 PEA-I P25, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA_1 P25, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence CKYCIIHQSTKLF in HSCP2 PEA-1 P25.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSCP2 PEA-1 P33, comprising a first amino acid sequence being at least 90 % homologous to MKILILGIFLFLCSTPAWAKEKHYYIGIIETTWDYASDHGEKKLISVDTEHSNIYLQNGPD
RIGRLYKKALYLQYTDETFRTTIEKPVWLGFLGPIIKAETGDKVYVHLKNLASRPYTFHS
HSHIDAPKDIASGLIGPLIICKK corresponding to amino acids 1 - 202 of CERU HUMAN, which also corresponds to amino acids 1 - 202 of HSCP2 PEA_1 P33, and a second amino acid sequence being at least 70°l°, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GTSSPYCTCYMTKRQGQGSLSFKKKSSLLC corresponding to amino acids 203 - 232 of HSCP2 PEA_1 P33, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSCP2 PEA_1 P33, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GTSSPYCTCYMTKRQGQGSLSFKKKSSLLC in HSCP2_PEA-1 P33.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN_PEA_1 P5, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQL.RCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQtaPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EV V VAE VGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
T corresponding to amino acids 1 - 1525 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 1525 of HUMTEN PEA-1 P5, a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence TEPKPQLGTLIFSNITPKSFNMSWTTQAGLFAKIVINVSDAHSLHESQQFTVSGDAKQAH
ITGLVENTGYDVS VAGTTLAGDPTRPLTAFVI corresponding to amino acids 1526 - 1617 of HUMTEN PEA_1 P5, and a third amino acid sequence being at least 90 %
homologous to TEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTWDSGKLLDPQEFTLSGTQRKLE
LRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTADE
GVFDNFVLKIRDTKKQSEPLEITLLAPERTRDLTGLREATEYEIELYGISKGRRSQTVSAI
ATTAMGSPKEV IFSDITENSATV SWRAPTAQVESFRITYV PITGGTPSM VTV DGTKTQTR
LVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIATV
PSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGDK
AQALEVFCDMTSDGGG WIVFLRRKNGRENFYQNWKAYAAGFGDRREEFWLGLDNLN
KITAQGQYELRVDLRDHGETAFAVYDKFSVGDAKTRYKLKVEGYSGTAGDSMAYHN
GRSFSTFDKDTDSAITNCALSYKGAFWYRNCHRVNLMGRYGDNNHSQGVNWFHWKG
HEHSIQFAEMKLRPSNFRNLEGRRKRA corresponding to amino acids 1526 - 2201 of TENA HUMAN V1, which also corresponds to amino acids 1618 - 2293 of HUMTEN PEA_1 P5, wherein said first amino acid sequence, second amino acid sequence and third amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of HUMTEN PEA_1 P5, comprising an amino acid sequence being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence encoding for TEPKPQLGTLIFSNITPKSFNMSWTTQAGLFAKIVINVSDAHSLHESQQFTVSGDAKQAH
ITGLVENTGYDVSVAGTTLAGDPTRPLTAFVI, corresponding to HUMTEN PEA_1 PS.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_1 P6, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPV SARV ATYLPAPEGLKFKSIKETS V E VEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EVVVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTE corresponding to amino acids 1 - 1527 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 1527 of HUMTEN PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PKPQLGTLIFSNITPKSFNMSWTTQAGLFAKIVINVSDAHSLHESQQFTVSGDAKQAHIT
GLVENTGYDVSVAGTTLAGDPTRPLTAFVITGTQSEVLTCLTQREKEISHLKGKFNKNTI
FTANVYSLIFN corresponding to amino acids 1528 - 1658 of HUMTEN PEA-1 P6, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence PKPQLGTLIFSNITPKSFNMSWTTQAGLFAKIVINVSDAHSLHESQQFTVSGDAKQAHIT
GLVENTGYDVSVAGTTLAGDPTRPLTAFVITGTQSEVLTCLTQREKEISHLKGKFNKNTI
FTANVYSLIFN in HUMTEN PEA I P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P7, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPV VFNH VYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
IO VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
I S FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATiNAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
EVWAEVGWDALKLN WTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTVVDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVT corresponding to amino acids 1 - 1617 of TENA HUMAN V1, which also corresponds to amino acids 1 - 1617 of I-IUMTEN PEA-1 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence GISNQVSHLFLFLVPFCVICLPDRHDFNIFVHIPYLIHKCSLLFHLLPTLPLVICT
corresponding to amino acids 1618 - 1673 of HUMTEN PEA-1 P7, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA_1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GISNQVSHLFLFLVPFCVICLPDRHDFNIFVHIPYLIHKCSLLFHLLPTLPLVICT in HUMTEN PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P8, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTV SIYGVIQGYRTPVLSAEASTGETPNLG
EVWAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
T corresponding to amino acids 1 - 1525 of TENA HUMAN V l, which also corresponds to amino acids 1 - 1525 of HUMTEN PEA-1 P8, and a second amino acid sequence being at least 90 % homologous to TEAEPEVDNLLV SDATPDGFRLSWTADEGVFDNFVLKIRDTKKQSEPLEITLLAPERTRD
LTGLREATEYEIELYGISKGRRSQTVSAIATTAMGSPKEVIFSDITENSATVSWRAPTAQV
ESFRITYVPITGGTPSMVTVDGTKTQTRLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTA
LDGPSGLVTANITDSEALARWQPAIATVDSYVISYTGEKVPEITRTVSGNTVEYALTDLE
PATEYTLRIFAEKGPQKSSTITAKFTTDLDSPRDLTATEVQSETALLTWRPPRASVTGYL
LVYESVDGTVKEVIVGPDTTSYSLADLSPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYP
FPKDCSQAMLNGDTTSGLYTIYLNGDKAQALEVFCDMTSDGGGWIVFLRRKNGRENF
YQNWKAYAAGFGDRREEFWLGLDNLNKITAQGQYELRVDLRDHGETAFAVYDKFSV
GDAKTRYKLKVEGYSGTAGDSMAYHNGRSFSTFDKDTDSAITNCALSYKGAFWYRNC
HRVNLMGRYGDNNHSQGVNWFHWKGHEHSIQFAEMKLRPSNFRNLEGRRKRA
corresponding to amino acids 1617 - 2201 of TENA_HUMAN V 1, which also corresponds to amino acids 1526 - 2110 of HUMTEN PEA_1 P8, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimerie polypeptide encoding for an edge portion of HUMTEN PEA-1 P8, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise TT, having a structure as follows: a sequence starting from any of amino acid numbers 1525-x to 1525; and ending at any of amino acid numbers 1526+ ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P10, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCI-ILRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EVWAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVL corresponding to amino acids 1 - 1252 of ilo TENA HUMAN V 1, which also corresponds to amino acids I - 1252 of HUMTEN PEA-1 P10, and a second amino acid sequence being at least 90 %
homologous to TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKV EAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTVVDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTAD
IATTAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQT
RLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIAT
VDSYVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTD
LDSPRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADL
SPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGD
KAQALEVFCDMTSDGGGWIVFLRRKNGRENFYQNWKAYAAGFGDRREEFWLGLDNL
NKITAQGQYELRVDLRDHGETAFAVYDKFSVGDAKTRYKLKVEGYSGTAGDSMAYH
NGRSFSTFDKDTDSAITNCALSYKGAFWYRNCHRVNLMGRYGDNNHSQGVNWFHWK
GHEHSIQFAEMKLRPSNFRNLEGRRKRA corresponding to amino acids 1344 - 2201 of TENA HUMAN V1, which also corresponds to amino acids 1253 - 2110 of HUMTEN PEA-1 P10, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMTEN PEA-1 P10, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise LT, having a structure as follows: a sequence starting from any of amino acid numbers 1252-x to 1252; and ending at any of amino acid numbers 1253+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P13, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSV DLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPV SARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EVWAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEW
corresponding to amino acids 1 - 1343 ofTENA HUMAN V1, which also corresponds to amino acids 1 - 1343 of HUMTEN PEA_1 P13, and a second amino acid sequence being at least 90 % homologous to TAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQTRLV
KLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIATVDS
YVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTDLDS
PRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADLSPS
THYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGDKAQ
ALEVFCDMTSDGGGWIVFLRRKNGRENFYQNWKAYAAGFGDRREEFWLGLDNLNKIT
AQGQYELRVDLRDI-IGETAFAVYDKFSVGDAKTRYKLKVEGYSGTAGDSMAYHNGRS
FSTFDKDTDSAITNCALSYKGAFWYRNCHRVNLMGRYGDNNHSQGVNWFHWKGHEH
SIQFAEMKLRPSNFRNLEGRRKRA corresponding to amino acids 1708 - 2201 of TENA HUMAN V1, which also corresponds to amino acids 1344 - 1837 of HUMTEN PEA 1 P13, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMTEN PEA-1 P
13, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise VT, having a structure as follows: a sequence starting from any of amino acid numbers 1343-x to 1343; and ending at any of amino acid numbers 1344+ ((rr2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_I P14, comprising a first amino acid sequence being at least 90 % homologous to LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTG LAPGQEYEISLHI V KNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EVVVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFI VYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTV SWMASENAFDSFLVTV VDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTAD
EGVFDNFVLKIRDTKKQSEPLEITLLAPERTRDLTGLREATEYEIELYGISKGRRSQTVSA
IATTAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQT
RLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIAT
VDSYVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTD
LDSPRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADL
SPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGD
KAQALEVFCDMTSDGGGWIV corresponding to amino acids 1 - 2025 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 2025 of HUMTEN PEA-1 P14, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence STTRDCRALRPRGRGRGQSRGGEEGDLLLMHSDTPMCEALQDSACHTEALRNSLLNKR
MGNTLATF corresponding to amino acids 2026 - 2091 of HUMTEN PEA_1 P14, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA_1 P 14, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence STTRDCRALRPRGRGRGQSRGGEEGDLLLMHSDTPMCEALQDSACHTEALRNSLLNKR
MGNTLATF in HUMTEN PEA 1 P14.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN_PEA_1 P15, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKAS corresponding to amino acids 1 - 1070 of TENA_HUMAN V 1, which also corresponds to amino acids 1 - 1070 of HUMTEN PEA-1 P15, and a second amino acid sequence being at least 90 % homologous to TEAEPEVDNLLVSDATPDGFRLSWTADEGVFDNFVLKIRDTKKQSEPLEITLLAPERTRD
LTGLREATEYEIELYGISKGRRSQTVSAIATTAMGSPKEVIFSDITENSATVSWRAPTAQV
ESFRITYVPITGGTPSMVTVDGTKTQTRLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTA
LDGPSGLVTANITDSEALARWQPAIATVDSYVISYTGEKVPEITRTVSGNTVEYALTDLE
PATEYTLRIFAEKGPQKSSTITAKFTTDLDSPRDLTATEVQSETALLTWRPPRASVTGYL
LVYESVDGTVKEVIVGPDTTSYSLADLSPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYP
FPKDCSQAMLNGDTTSGLYTIYLNGDKAQALEVFCDMTSDGGGWIVFLRRKNGRENF
YQNWKAYAAGFGDRREEFWLGLDNLNKITAQGQYELRVDLRDHGETAFAVYDKFSV
GDAKTRYKLKVEGYSGTAGDSMAYHNGRSFSTFDKDTDSAITNCALSYKGAFWYRNC
HRVNLMGRYGDNNHSQGVNWFHWKGHEHSIQFAEMKLRPSNFRNLEGRRKRA
corresponding to amino acids 1617 - 2201 of TENA HUMAN V 1, which also corresponds to amino acids 1071 - 1655 of HUMTEN PEA-1 P15, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMTEN PEA-1 P15, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise ST, having a structure as follows: a sequence starting from any of amino acid numbers 1070-x to 1070; and ending at any of amino acid numbers 1071+ ((m2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_I P16, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
FTGPDCAELSCPNDCI-IGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLV VTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPS.QIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRV SQTDNSITLEWRNGKAAIDSYRI KYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKAS corresponding to amino acids I - 1070 of TENA HUMAN VI, which also corresponds to amino acids I - 1070 of HUMTEN PEA-1 P16, and a second amino acid sequence being at least 90 % homologous to TAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQTRLV
KLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIATVDS
YVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTDLDS
PRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADLSPS
THYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGDKAQ
ALEVFCDMTSDGGGWIVFLRRKNGRENFYQNWKAYAAGFGDRREEFWLGLDNLNKIT
AQGQYELRVDLRDHGETAFAVYDKFSVGDAKTRYKLKVEGYSGTAGDSMAYHNGRS
FSTFDKDTDSAITNCALSYKGAFWYRNCHRVNLMGRYGDNNHSQGVNWFHWKGHEH
SIQFAEMKLRPSNFRNLEGRRKRA corresponding to amino acids 1708 - 2201 of TENA_HUMAN V 1, which also corresponds to amino acids 1071 - 1564 of HUMTEN PEA_1 P16, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMTEN PEA_1 P16, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise ST, having a structure as follows: a sequence starting from any of amino acid numbers 1070-x to 1070; and ending at any of amino acid numbers 107 I+ ((rr2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P 17, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDP LDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLR.AVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EV VVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTP LSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTVVDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTAD
EGVFDNFVLKIRDTKKQSEPLEITLLAPERTRDLTGLREATEYEIELYGISKGRRSQTVSA
IATTAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQT
RLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIAT
VDSYVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTD
LDSPRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADL
SPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGD
KAQALEVFCDMTSDGGGWIV corresponding to amino acids 1 - 2025 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 2025 of HUMTEN PEA-1 P17, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence TPWPTTMADPSPPLTRTQIQPSPTVLCPTKGLSGTGTVTVST corresponding to amino acids 2026 - 2067 of HUMTEN PEA-1 P17, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA-1 P17, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TPWPTTMADPSPPLTRTQIQPSPTVLCPTKGLSGTGTVTVST in HUMTEN PEA 1 P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P20, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCI-INRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTI DLTEDENQYSIGNLKPDTEYEV SLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVK.EDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQW VGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTV SIYGVIQGYRTPVLSAEASTGETPNLG
EVVVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTVVDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTAD
EGVFDNFVLKIRDTKKQSEPLEITLLAPERTRDLTGLREATEYEIELYGISKGRRSQTVSA
IATTAMGSPKEVIFSDITENSATVSWRAPTAQVESFRITYVPITGGTPSMVTVDGTKTQT
RLVKLIPGVEYLVSIIAMKGFEESEPVSGSFTTALDGPSGLVTANITDSEALARWQPAIAT
VDSYVISYTGEKVPEITRTVSGNTVEYALTDLEPATEYTLRIFAEKGPQKSSTITAKFTTD
LDSPRDLTATEVQSETALLTWRPPRASVTGYLLVYESVDGTVKEVIVGPDTTSYSLADL
SPSTHYTAKIQALNGPLRSNMIQTIFTTIGLLYPFPKDCSQAMLNGDTTSGLYTIYLNGD
KAQALEVFCDMTSDGGGWIVFLRRKNGRENFYQNWKAYAAGFGDRREEFWLG
corresponding to amino acids 1 - 2057 of TENA HUMAN V1, which also corresponds to amino acids 1 - 2057 of HUMTEN_PEA-1 P20, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NAALHVYI
corresponding to amino acids 2058 - 2065 of I-IUMTEN PEA-l P20, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN_PEA_1 P20, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NAALHVYI in HUMTEN PEA 1 P20.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P26, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPWFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EVVVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGL.K
AATHYTITIRGVTQDFSTTPLSVEVLTEEVP DMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
TEDLPQLGDLAVSEVGWDGLRLNWTAADNAYEHFVIQVQEVNKVEAAQNLTLPGSLR
AVDIPGLEAATPYRVSIYGVIRGYRTPVLSAEASTAKEPEIGNLNVSDITPESFNLSWMA
TDGIFETFTIEIIDSNRLLETVEYNISGAERTAHISGLPPSTDFIVYLSGLAPSIRTKTISATA
TTEALPLLENLTISDINPYGFTVSWMASENAFDSFLVTVVDSGKLLDPQEFTLSGTQRKL
ELRGLITGIGYEVMVSGFTQGHQTKPLRAEIVTEAEPEVDNLLVSDATPDGFRLSWTAD
EGVFDNFVLKIRDTKKQSEPLEITLLAPERTRDLTGLREATEYEIELYGISKGRRSQTVSA
IATT corresponding to amino acids 1 - 1708 of TENA HUMAN V1, which also corresponds to amino acids 1 - 1708 of HUMTEN PEA-1 P26, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence 1 S GTVNKQERTEKSHDSGVFFSQG corresponding to amino acids 1709 - 1730 of HUMTEN PEA-1 P26, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA 1 P26, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GTVNKQERTEKSHDSGVFFSQG in HUMTEN PEA_1 P26.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_1 P27, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLV VYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRA VDIPGLKAATPYTV SIYGVIQGYRTPVLSAEASTGETPNLG
EVWAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLTEEVPDMGNLTVTEVSWDALRLNWTTPDGTYD
QFTIQVQEADQVEEAHNLTVPGSLRSMEIPGLRAGTPYTVTLHGEVRGHSTRPLAVEVV
T corresponding to amino acids 1 - 1344 of TENA HUMAN V1, which also corresponds to amino acids 1 - 1344 of HUMTEN PEA-1 P27, and a second amino acid sequence being at least.70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GI
corresponding to amino acids 1345 - 1346 of HUMTEN PEA-1 P27, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P28, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLV VYTPTHEGGLEMQFRVPGDQTSTIIQELEPGV EYFIRVFAI
LENKKSIPV SARVATYLPAPEGLKFKSIKETSVEVEW DPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRV SQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKASTEQAPELENLTVTEVGWDGLRLNWTAADQAYEHFIIQVQE
ANKVEAARNLTVPGSLRAVDIPGLKAATPYTVSIYGVIQGYRTPVLSAEASTGETPNLG
EWVAEVGWDALKLNWTAPEGAYEYFFIQVQEADTVEAAQNLTVPGGLRSTDLPGLK
AATHYTITIRGVTQDFSTTPLSVEVLT corresponding to amino acids 1 - 1253 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 1253 of HUMTEN PEA-1 P28, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GILDEFTNSLPPLCLCSGGIKALSCFKLGSAPTTLGKYQ corresponding to amino acids 1254 - 1292 of HUMTEN PEA-I P28, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA-1 P28, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GILDEFTNSLPPLCLCSGGIKALSCFKLGSAPTTLGKYQ in HUMTEN PEA_1 P28.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_1 P29, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRI-IKRQSGVNATLPEENQPVVFNHVYNIK
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVC.DEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTGLRPGTEYGIGVSAVKEDKESNPATINAATELDTPKDLQVSE
TAETSLTLLWKTPLAKFDRYRLNYSLPTGQWVGVQLPRNTTSYVLRGLEPGQEYNVLL
TAEKGRHKSKPARVKAST corresponding to amino acids 1 - 1071 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 1071 of HUMTEN PEA-1 P29, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GESALSFLQTLG corresponding to amino acids 1072 - 1083 of HUMTEN PEA-1 P29, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA-1 P29, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GESALSFLQTLG in HUMTEN PEA 1 P29.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_1 P30, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTRLDAPSQIEVKDVT
DTTALITWFKPLAEIDGIELTYGIKDVPGDRTTIDLTEDENQYSIGNLKPDTEYEVSLISRR
GDMSSNPAKETFTTGLDAPRNLRRVSQTDNSITLEWRNGKAAIDSYRIKYAPISGGDHA
EVDVPKSQQATTKTTLTG corresponding to amino acids 1 - 954 of TENA HUMAN V l, which also corresponds to amino acids 1 - 954 of HUMTEN PEA-1 P30, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence ELCISASLSQPALEGP corresponding to amino acids 955 - 970 of HUMTEN PEA-1 P30, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA-1 P30, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence ELCISASLSQPALEGP in HUMTEN PEA 1 P30.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA_1_P31, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCI-ILRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLWTEVTE
I S ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVATYLPAPEGLKFKSIKETSVEVEWDPLDIAFETWEIIFRNMNKEDEG
EITKSLRRPETSYRQTGLAPGQEYEISLHIVKNNTRGPGLKRVTTTR corresponding to amino acids 1 - 802 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 802 of HUMTEN PEA_1 P31, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence EYHL corresponding to amino acids 803 - 806 of HUMTEN PEA-1 P31, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMTEN PEA_1 P31, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% ho mologous to the sequence EYHL
in HUMTEN PEA 1 P31.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMTEN PEA-1 P32, comprising a first amino acid sequence being at least 90 % homologous to MGAMTQLLAGVFLAFLALATEGGVLKKVIRHKRQSGVNATLPEENQPVVFNHVYNIK
LPVGSQCSVDLESASGEKDLAPPSEPSESFQEHTVDGENQIVFTHRINIPRRACGCAAAP
DVKELLSRLEELENLVSSLREQCTAGAGCCLQPATGRLDTRPFCSGRGNFSTEGCGCVC
EPGWKGPNCSEPECPGNCHLRGRCIDGQCICDDGFTGEDCSQLACPSDCNDQGKCVNG
VCICFEGYAGADCSREICPVPCSEEHGTCVDGLCVCHDGFAGDDCNKPLCLNNCYNRG
RCVENECVCDEGFTGEDCSELICPNDCFDRGRCINGTCYCEEGFTGEDCGKPTCPHACH
TQGRCEEGQCVCDEGFAGVDCSEKRCPADCHNRGRCVDGRCECDDGFTGADCGELKC
PNGCSGHGRCVNGQCVCDEGYTGEDCSQLRCPNDCHSRGRCVEGKCVCEQGFKGYDC
SDMSCPNDCHQHGRCVNGMCVCDDGYTGEDCRDRQCPRDCSNRGLCVDGQCVCEDG
FTGPDCAELSCPNDCHGRGRCVNGQCVCHEGFMGKDCKEQRCPSDCHGQGRCVDGQ
CICHEGFTGLDCGQHSCPSDCNNLGQCVSGRCICNEGYSGEDCSEVSPPKDLVVTEVTE
ETVNLAWDNEMRVTEYLVVYTPTHEGGLEMQFRVPGDQTSTIIQELEPGVEYFIRVFAI
LENKKSIPVSARVAT corresponding to amino acids 1 - 710 of TENA HUMAN V 1, which also corresponds to amino acids 1 - 710 of HUMTEN PEA-I P32, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence CE corresponding to amino acids 711 - 712 of HUMTEN PEA-1 P32, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMOSTRO PEA-1 PEA-1 P21, comprising a first amino acid sequence being at least 90 % homologous to MRIAVICFCLLGITCAIPVKQADSGSSEEKQLYNKYPDAVATWLNPDPSQKQNLLAPQ
corresponding to amino acids 1 - 58 of OSTP HUMAN, which also corresponds to amino acids 1 - 58 of HUMOSTRO PEA-1 PEA-1 P21, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VFLNFS
corresponding to amino acids 59 - 64 of HUMOSTRO PEA_1 PEA-1 P21, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMOSTRO PEA 1 PEA-1 P21, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence VFLNFS in HUMOSTRO PEA 1 PEA 1 P21.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMOSTRO PEA-1 PEA-1 P25, comprising a first amino acid sequence being at least 90 % homologous to MRIAVICFCLLGITCAIPVKQADSGSSEEKQ corresponding to amino acids 1 - 31 of OSTP HUMAN, which also con-esponds to amino acids 1 - 3 l of HUMOSTRO PEA-1 PEA-1 P25, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence H
corresponding to amino acids 32 - 32 of HUMOSTRO PEA_1 PEA-1 P25, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMOSTRO PEA-1 PEA_1 P30, comprising a first amino acid sequence being at least 90 % homologous to MRIAVICFCLLGITCAIPVKQADSGSSEEKQ corresponding to amino acids 1 - 31 of OSTP HUMAN, which also corresponds to amino acids 1 - 31 of HUMOSTRO PEA_1 PEA-1 P30, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence VSIFYVFI
corresponding to amino acids 32 - 39 of HUMOSTRO PEA-1 PEA-1 P30, wherein said first amino acid sequence and second amino acid sequence are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMOSTRO PEA_1 PEA 1 P30, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence VSIFYVFI in HUMOSTRO PEA 1 PEA 1 P30.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H61775 P16, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPR1DPDYVG corresponding to amino acids 1 t - 93 of Q9P2J2, which also corresponds to amino acids 1 - 83 of I-161775 P16, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV corresponding to amino acids 84 - 152 of H61775 P16, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of H61775 P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV in H61775 P16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H61775 P 16, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSWGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 1 - 83 of AAQ88495, which also corresponds to amino acids 1 - 83 of H61775 P16, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV corresponding to amino acids 84 - 152 of H61775 P16, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of H61775 P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV in H61775 P16.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H61775 P17, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 1 I - 93 of Q9P2J2, which also corresponds to amino acids 1 - 83 of H61775_P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H61775 P17, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 1 - 83 of AAQ88495, which also corresponds to amino acids 1 - 83 of H61775 P17.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P2, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PHSGPAAAFIRRRGWWPGPRCA corresponding to amino acids 1 - 22 of HSAPHOL P2, second amino acid sequence being at least 90 % homologous to PATPRPLSWLRAPTRLCLDGPSPVLCA corresponding to amino acids 1 - 27 of AAH21289, which also corresponds to amino acids 23 - 49 of HSAPHOL P2, and a third amino acid sequence being at least 90 % homologous to EKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFLGDGMGVSTVTAARILKGQL
HHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLCGVKANEGTVGVSAAT
ERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAAYAHSADRDWYSDNE
MPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDVEYESDEKARGTRLD
GLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEPGDMQYELNRNNVT
DPSLSEM VWAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALHEAVEMDRAIGQAG
SLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYK
VVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQN
YVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 83 - 586 of AAH21289, which also corresponds to amino acids 50 -553 of I-ISAPHOL P2, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of HSAPHOL P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence PHSGPAAAFIRRRGWWPGPRCA of HSAPHOL P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of I-ISAPHOL P2, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise AE, having a structure as follows: a sequence starting from any of amino acid numbers 49-x to 50; and ending at any of amino acid numbers 50+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL_P2, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PHSGPAAAFIRRRGWWPGPRCAPATPRPLSWLRAPTRLCLDGPSPVLCA corresponding to amino acids 1 - 49 of HSAPHOL P2, second amino acid sequence being at least homologous to EKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFLGDGMGVSTVTAARILKGQL
HHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLCGVKANEGTVGVSAAT
ERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAAYAHSADRDWYSDNE
MPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDVEYESDEKARGTRLD
GLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEPGDMQYELNRNNVT
DPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALHEAVEMDRAIGQAG
SLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYK
WGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQN
YVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 21 - 524 of PPBT HUMAN, which also corresponds to amino acids 50 -553 of HSAPE-lOL-P2, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of HSAPHOL P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence PHSGPAAAFIRRRGWWPGPRCAPATPRPLSWLRAPTRLCLDGPSPVLCA of HSAPHOL P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P2, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise AE, having a structure as follows: a sequence starting from any of amino acid numbers 49-x to S0; and ending at any of amino acid numbers 50+ ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P3, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVP corresponding to amino acids 63 - 82 of AAH21289, which also corresponds to amino acids 1 - 20 of HSAPHOL P3, and a second amino acid sequence being at least 90 % homologous to GMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYL
CGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSA
AYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTD
VEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFE
PGDMQYELNRNNVTDPSLSEMV VVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQAL
HEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKK
PFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKG
PMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSV
LF corresponding to amino acids 123 - 586 of AAH21289, which also corresponds to amino acids 21 - 484 of HSAPHOL P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P3, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise PG, having a structure as follows: a sequence starting from any of amino acid numbers 20-x to 20; and ending at any of amino acid numbers 21+ ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P3, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVP corresponding to amino acids 1 - 20 of PPBT HUMAN, which also corresponds to amino acids 1 - 20 of HSAPHOL P3, and a second amino acid sequence being at least 90 % homologous to GMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYL
CGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSA
AYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTD
VEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFE
PGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQAL
HEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKK
PFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKG
PMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSV
LF corresponding to amino acids 61 - 524 of PPBT HUMAN, which also corresponds to amino acids 21 - 484 of HSAPHOL P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P3, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise PG, having a structure as follows: a sequence starting from any of amino acid numbers 20-x to 20; and ending at any of amino acid numbers 21+ ((n-2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P4, comprising a first amino acid sequence being at least 90 % homologous to MGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLC
GVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAA
YAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDV
EYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEP
GDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALH
EAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKP
FTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGP
MAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVL
F corresponding to amino acids 124 - 586 of AAH21289, which also corresponds to amino acids 1 - 463 of HSAPHOL P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P4, comprising a first amino acid sequence being at least 90 % homologous to MGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLC
GVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAA
YAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDV
EYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEP
GDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALH
EAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKP
FTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGP
MAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVL
F corresponding to amino acids 62 - 524 of PPBT HUMAN, which also corresponds to amino acids 1 - 463 of HSAPHOL P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P5, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
GLFEPGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKA
KQALHEAVEM corresponding to amino acids 63 - 417 of AAH21289, which also corresponds to amino acids 1 - 355 of HSAPHOL P5, and a second amino acid sequence being at least 90 homologous to DHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVD
YAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIG
ANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 440 - 586 of AAH21289, which also corresponds to amino acids 356 - 502 of HSAPHOL PS, wherein said 1 S first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P5, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise MD, having a structure as follows: a sequence starting from any ofamino acid numbers 355-x to 355; and ending at any of amino acid numbers 356+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHO L P5, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
GLFEPGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHI-IEGKA
KQALHEAVEM corresponding to amino acids 1 - 355 of .PPBT HUMAN, which also corresponds to amino acids 1 - 355 of HSAPHOL_P5, and a second amino acid sequence being at least 90 % homologous to DHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVD
YAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIG
ANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 377 - 524 of PPBT HUMAN, which also corresponds to amino acids 356 - 502 of HSAPHOL P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P5, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise MD, having a structure as follows: a sequence starting from any of amino acid numbers 355-x to 355; and ending at any of amino acid numbers 356+ ((rr2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P6, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
corresponding to amino acids 63 - 349 of AAH21289, which also corresponds to amino acids 1 287 of HSAPHOL P6, and a second amino acid sequence being at least 90 %
homologous to GGRIDHGHHEGKAKQALHEAVEMDRAIGQAGSLTSSEDTLTWTADHSHVFTFGGYTP
RGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAV
PLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAG
SLAAGPLLLALALYPLSVLF corresponding to amino acids 395 - 586 of AAH21289, which also corresponds to amino acids 288 - 479 of i-1SAPHOL P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P6, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise LG, having a structure as follows: a sequence starting from any of amino acid numbers 287-x to 287; and ending at any of amino acid numbers 288+ ((rr2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P6, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
corresponding to amino acids 1 - 287 of PPBT HUMAN, which also corresponds to amino acids 1 - 287 of HSAPHOL P6, and a second amino acid sequence being at least homologous to GGRIDHGHHEGKAKQALHEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTP
RGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAV
PLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAG
SLAAGPLLLALALYPLSVLF corresponding to amino acids 333 - 524 of PPBT HUMAN, which also corresponds to amino acids 288 - 479 of HSAPHOL Pti, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P6, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise LG, having a structure as follows: a sequence starting from any of amino acid numbers 287-x to 287; and ending at any of amino acid numbers 288+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P7, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYK corresponding to amino acids 63 -326 of AAH21289, which also corresponds to amino acids 1 - 264 of HSAPHOL P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, 1 S more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence LPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP
corresponding to amino acids 265 - 306 of HSAPHOL P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence LPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP in HSAPHOL P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P7, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPR corresponding to amino acids 1 - 262 of PPBT HUMAN, which also corresponds to amino acids 1 - 262 of HSAPHOL-P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence YKLPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP
corresponding to amino acids 263 - 306 of I-ISAPHOL P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence YKLPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP in HSAPHOL P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P7, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYK corresponding to amino acids 1 -264 of 075090, which also corresponds to amino acids 1 - 264 of HSAPHOL P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence LPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP corresponding to amino acids 265 - 306 of HSAPHOL P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence LPPRCPLANRVDFSWAGREYRLQTFSKPLIFLANVFLQTQRP in HSAPHOL P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P8, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKI-ISHFIWNRTELLTLDPHNVDYLL
G corresponding to amino acids 63 - 350 of AAH21289, which also corresponds to amino acids 1 - 288 of HSAPHOL P8, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP corresponding to amino acids 289 - 316 of HSAPHOL P8, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P8, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP in HSAPHOL P8.
According to preferred embodiments of the present invention, there is provided an isolated chimerie polypeptide encoding for HSAPHOL P8, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
G corresponding to amino acids 1 - 288 of PPBT HUMAN, which also corresponds to amino acids 1 - 288 of HSAPHOL P8, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP corresponding to amino acids 289 - 316 of HSAPHOL P8, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P8, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP in HSAPHOL P8.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSAPHOL P8, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
G corresponding to amino acids 1 - 288 of 075090, which also corresponds to amino acids 1 -288 of HSAPHOL P8, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP corresponding to amino acids 289 - 316 of HSAPHOL P8, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSAPHOL P8, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KWRGWRGGCMARSLVAGAACGQHLGTRP in HSAPHOL P8.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T10888 PEA_1 P2, comprising a first amino acid sequence being at least 90 % homologous to MGPPSAPPCRLHVPWKEVLLTASLLTFWNPPTTAKLTIESTPFNVAEGKEVLLLAHNLP
QNRIGYSWYKGERVDGNSLIVGYVIGTQQATPGPAYSGRETIYPNASLLIQNVTQNDTG
FYTLQVIKSDLVNEEATGQFHVYPELPKPSISSNNSNPVEDKDAVAFTCEPEVQNTTYL
WWVNGQSLPVSPRLQLSNGNMTLTLLSVKRNDAGSYECEIQNPASANRSDPVTLNVLY
YMCQAHNSATGLNRTTVTMITVS corresponding to amino acids 1 - 319 of CEA6 HUMAN, which also corresponds to amino acids 1 - 319 of T10888 PEA_1 P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DWTRP corresponding to amino acids 320 - 324 of T10888 PEA_1 P2, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T10888 PEA_I P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DWTRP in T10888 PEA 1 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T10888 PEA_I P4, comprising a first amino acid sequence being at least 90 % homologous to MGPPSAPPCRLHVPWKEVLLTASLLTFWNPPTTAKLTIESTPFNVAEGKEVLLLAHNLP
QNRIGYSWYKGERVDGNSLIVGYVIGTQQATPGPAYSGRETIYPNASLLIQNVTQNDTG
WWVNGQSLPVSPRLQLSNGNMTLTLLSVKRNDAGSYECEIQNPASANRSDPVTLNVL
corresponding to amino acids 1 - 234 of GEA6 HUMAN, which also corresponds to amino acids 1 - 234 of T10888 PEA I P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence LLLSSQLWPPSASRLECWPGWL corresponding to amino acids 235 - 256 of T10888 PEA_1 P4, wherein said first and second amino acid sequences are contiguois and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T10888 PEA-1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence LLLSSQLWPPSASRLECWPGWL in T10888 PEA 1 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T10888 PEA-1 P4, comprising a first amino acid sequence being at least 90 % homologous to MGPPSAPPCRLHVPWKEVLLTASLLTFWNPPTTAKLTIESTPFNVAEGKEVLLLAHNLP
QNRIGYSWYKGERVDGNSLIVGYVIGTQQATPGPAYSGRETIYPNASLLIQNVTQNDTG
FYTLQVIKSDLVNEEATGQFHVYPELPKPSISSNNSNPVEDKDAVAFTCEPEVQNTTYL
W WVNGQSLPVSPRLQLSNGNMTLTLLSVKRNDAGSYECEIQNPASANRSDPVTLNVL
corresponding to amino acids 1 - 234 of Q13774, which also corresponds to amino acids 1 - 234 of T10888 PEA-1 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence LLLSSQLWPPSASRLECWPGWL
corresponding to amino acids 235 - 256 of T10888 PEA_1 P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T10888 PEA-1 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence LLLSSQLWPPSASRLECWPGWL in T10888 PEA 1 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T10888 PEA-1 P5, comprising a first amino acid sequence being at least 90 % homologous to MGPPSAPPCRLHVPWKEVLLTASLLTFWNPPTTAKLTIESTPFNVAEGKEVLLLAHNLP
QNRIGYSWYKGERVDGNSLIVGYVIGTQQATPGPAYSGRETIYPNASLLIQNVTQNDTG
FYTLQVIKSDLVNEEATGQFHVYPELPKPSISSNNSNPVEDKDAVAFTCEPEVQNTTYL
WWVNGQSLPVSPRLQLSNGNMTLTLLSVKRNDAGSYECEIQNPASANRSDPVTLNVLY
GPDVPTISPSKANYRPGENLNLSCHAASNPPAQYSWFINGTFQQSTQELFIPNITVNNSGS
YMCQAHNSATGLNRTTVTMITVSG corresponding to amino acids 1 - 320 of CEA6 HUMAN, which also corresponds to amino acids 1 - 320 of T 10888 PEA-1 P5, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide havingthe sequence VVFCFLISHV corresponding to amino acids 321 - 390 of T10888 PEA-1 P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T10888 PEA-1 P5, comprising a polypeptide being at. least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KWIHEALASHFQVESGSQRRARKKFSFPTCVQGAHANPKFSPEPSQFTSADSFPLVFLFF
WFCFLISHV in T10888 PEA 1 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T10888 PEA-1 P6, comprising a first amino acid sequence being at least 90 % homologous to MGPPSAPPCRLHVPWKEVLLTASLLTFWNPPTTAKLTIESTPFNVAEGKEVLLLAHNLP
QNRIGYSWYKGERVDGNSLIVGYVIGTQQATPGPAYSGRETIYPNASLLIQNVTQNDTG
FYTLQVIKSDLVNEEATGQFHW
corresponding to amino acids 1 - 141 of CEA6 HUMAN, which also corresponds to amino acids 1 - 141 of T10888 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence REYFHMTSGCWGSVLLPTYGIVRPGLCLWPSLHYILYQGLDI
corresponding to amino acids 142 - 183 of T10888 PEA 1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T10888 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence REYFHMTSGCWGSVLLPTYGIVRPGLCLWPSLHYILYQGLDI in T10888 PEA_I P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for I-ISECADH P9, comprising a first amino acid sequence being at least 90 % homologous to CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASV SGIQAELLTFPNSSPGLRRQKRDW VIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEG corresponding to amino acids 1 -274 of Q9UII7, which also corresponds to amino acids 1 - 274 of HSECADH P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG
corresponding to amino acids 275 - 322 of HSECADH P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSECADH_P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG in HSECADH P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH_P9, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEG corresponding to amino acids I -274 of Q9UII8, which also corresponds to amino acids 1 - 274 of HSECADH P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG
corresponding to amino acids 275 - 322 of HSECADH P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of I-ISECADH P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG in HSECADH P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P9, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEG corresponding to amino acids 1 -274 of CAD1 HUMAN, which also corresponds to amino acids 1 - 274 of HSECADH
P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homobgous to a polypeptide having the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG corresponding to amino acids 275 - 322 of HSECADH P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSECADH P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TACRSRIANSCHSGDSWRNSCFANSDSAALAVSSEESGGQRALTAPRG in HSECADH P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P13, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFTINRNTGVISVVTTGLDRESFPTYTLVVQAADLQGEGL
STTATAVITVTDTNDNPPIFNPTT corresponding to amino acids 1 - 379 of Q9UII7, which also corresponds to amino acids 1 - 379 of HSECADH P13, and a second amino acid sequence VIL corresponding to amino acids 380 - 382 of HSECADH P13, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P 13, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFTINRNTGVISVVTTGLDRESFPTYTLVVQAADLQGEGL
STTATAVITVTDTNDNPPIFNPTT corresponding to amino acids 1 - 379 of Q9UII8, which also corresponds to amino acids I - 379 of HSECADH P13, and a second amino acid sequence VIL corresponding to amino acids 380 - 382 of HSECADH P13, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P13, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRI-ILERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
S GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFTINRNTGVISVVTTGLDRESFPTYTLVVQAADLQGEGL
STTATAV1TVTDTNDNPPIFNPTT corresponding to amino acids 1 - 379 of CAD 1-HUMAN, which also corresponds to amino acids 1 - 379 ofHSECADH_P13, and a second amino acid sequence V1L corresponding to amino acids 380 - 382 of HSECADH P13, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P 14, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAV SSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFTINRNTGVISVVTTGLDRE corresponding to amino acids 1 - 336 of Q9UII7, which also corresponds to amino acids 1 - 336 of HSECADH P
14, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRGQEDPEGVEDKCVLAQSRGQSK.ILLGQLSVNTVMV
corresponding to amino acids 337 - 373 of HSECADH P14, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSECADH P14, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRGQEDPEGVEDKCVLAQSRGQSKILLGQLSVNTVMV in HSECADH P14.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P 14, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFT1NRNTGVISVVTTGLDRE corresponding to amino acids 1'- 336 of Q9UII8, which also corresponds to amino acids 1 - 336 of HSECADH
P14, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 94% and most preferably at least 95% homologous to a polypeptide having the sequence VRGQEDPEGVEDKCVLAQSRGQSKILLGQLSVNTVMV
corresponding to amino acids 337 - 373 of HSECADH P14, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSECADH P 14, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRGQEDPEGVEDKCVLAQSRGQSKILLGQLSVNTVMV in HSECADH P14.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P14, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSN
GNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNT
YNAAIAYTILSQDPELPDKNMFTINRNTGVISWTTGLDRE corresponding to amino acids 1 - 336 of CADI HUMAN, which also corresponds to amino acids I - 336 of HSECADH P14, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRGQEDPEGVEDKCVLAQSRGQSKILLGQLSVNTVMV
corresponding to amino acids 337 - 373 of HSECADH P14, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HSECADH P14, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRGQEDPEGVEDKCVLAQSRGQSKILLGQLSVNTVMV in HSECADH P14.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for I-ISECADH P15, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASV SGIQAELLTFPNSSPGLRRQKRDW VIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYT corresponding to amino acids 1 - 229 of Q9UII7, which also corresponds to amino acids 1 -229 of HSECADH P15, and a second amino acid sequence VSIS corresponding to amino acids 230 -233 of HSECADH P15, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P15, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYT corresponding to amino acids 1 - 229 of Q9UII8, which also corresponds to amino acids 1 -229 of HSECADH P15, and a second amino acid sequence VSIS corresponding to amino acids 230 -233 of HSECADH P15, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSECADH P15, comprising a first amino acid sequence being at least 90 % homologous to MGPWSRSLSALLLLLQVSSWLCQEPEPCHPGFDAESYTFTVPRRHLERGRVLGRVNFED
CTGRQRTAYFSLDTRFKVGTDGVITVKRPLRFHNPQIHFLVYAWDSTYRKFSTKVTLNT
VGHHHRPPPHQASVSGIQAELLTFPNSSPGLRRQKRDWVIPPISCPENEKGPFPKNLVQI
KSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYT corresponding to amino acids 1 - 229 of CAD1 HUMAN, which also corresponds to amino acids 1 -229 of HSECADH P15, and a second amino acid sequence VSIS corresponding to amino acids 230 -233 of HSECADH P15, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P5, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYK corresponding to amino acids 12 - 55 of GILT HUMAN, which also corresponds to amino acids 1 - 44 of T59832 P5, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VGTATGRAGWREQAPCRGTRLLLSPQTSQGKTRAPRGRCPCRVPGKTLFSSRRCGHTP
SVPFRFRIPHLRGAAASTRLVPPKGSMSAYCVLLGQELGSPFVAQGTSSAAGQGPPACIL
AATLDAFIPARAGLACLWDLLGRCPRG corresponding to amino acids 45 - 189 of T59832 P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P5, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VGTATGRAGWREQAPCRGTRLLLSPQTSQGKTRAPRGRCPCRVPGKTLFSSRRCGHTP
SVPFRFRIPHLRGAAASTRLVPPKGSMSAYCVLLGQELGSPFVAQGTSSAAGQGPPACIL
AATLDAFIPARAGLACLWDLLGRCPRG in T59832 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P7, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQ ALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNG corresponding to amino acids 12 - 223 of GILT HUMAN, which also corresponds to amino acids 1 - 212 of T59832 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRIFLALSLTLIVPWSQGWTRQRDQR corresponding to amino acids 213 - 238 of T59832 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRIFLALSLTLIVPWSQGWTRQRDQR in T59832 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P7, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNG corresponding to amino acids 1 - 212 of BAC98466, which also corresponds to amino acids 1 - 212 of T59832 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRIFLALSLTLIVPWSQGWTRQRDQR corresponding to amino acids 213 - 238 of T59832 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRIFLALSLTLIVPWSQGWTRQRDQR in T59832 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P7, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV corresponding to amino acids 1 - 90 of T59832 P7, and a second amino acid sequence being at least 90 % homologous to MEILNVTLVPYGNAQEQNVSGRWEFKCQHGEEECKFNKVEACVLDELDMELAFLTIVC
MEEFEDMERSLPLCLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYV
PWVTVNGVRIFLALSLTLIVPWSQGWTRQRDQR corresponding to amino acids 1 - 148 of BAC85622, which also corresponds to amino acids 91 - 238 of T59832_P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of T59832 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV of T59832 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P7, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNG corresponding to amino acids 1 - 212 of Q8WU77, which also corresponds to amino acids 1 - 212 of.T59832 P7, and a second amino acid sequence being at least 70~%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VRIFLALSLTLIVPWSQGWTRQRDQR corresponding to amino acids 213 - 238 of T59832 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VRIFLALSLTLIVPWSQGWTRQRDQR in T59832 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P9, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHE corresponding to amino acids 12 - 214 of GILT HUMAN, which also corresponds to amino acids 1 - 203 of T59832 P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR corresponding to amino acids 204 - 244 of T59832 P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR in T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P9, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHE corresponding to amino acids 1 - 203 of BAC98466, which also corresponds to amino acids 1 - 203 of T59832 P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR corresponding to amino acids 204 - 244 of T59832 P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR in T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P9, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV corresponding to amino acids 1 - 90 of T59832 P9, second amino acid sequence being at least 90 % homologous to MEILNVTLVPYGNAQEQNVSGRWEFKCQHGEEECKFNKVEACVLDELDMELAFLTIVC
MEEFEDMERSLPLCLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHE
corresponding to amino acids 1 - 113 of BAC85622, which also corresponds to amino acids 91 -203 of T59832 P9, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR corresponding to amino acids 204 - 244 of T59832 P9, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV of T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR in T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P9, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVEACVLDELDMELAFLTIVCMEEFEDMERSLPLCLQLYAPGLSPDTIM
ECAMGDRGMQLMHANAQRTDALQPPHE corresponding to amino acids 1 - 203 of Q8WU77, which also corresponds to amino acids 1 - 203 of T59832 P9, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR corresponding to amino acids 204 - 244 of T59832 P9, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P9, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NPWKIRPSSLPLSASCTRARSRMSALPQPAPSGVFASSDGR in T59832 P9.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P12, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVE corresponding to amino acids 12 - 141 of GILT HUMAN, which also corresponds to amino acids 1 - 130 of T59832 P12, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 173 - 261 of GILT HUMAN, which also corresponds to amino acids 131 - 219 of T59832 P12, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P12, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EC, having a structure as follows: a sequence starting from any of amino acid numbers 130-x to 130; and ending at any of amino acid numbers 131+ ((n-2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P12, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV corresponding to amino acids 1 - 90 of T59832 P12, second amino acid sequence being at least 90 % homologous to MEILNVTLVPYGNAQEQNVSGRWEFKCQHGEEECKFNKVE corresponding to amino acids 1 - 40 of BAC85622, which also corresponds to amino acids 91 - 130 of T59832 P 12, third amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNG
corresponding to amino acids 72 - 122 of BAC85622, which also corresponds to amino acids 131 - 181 of T59832 P12, and a fourth amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence KPLEDQTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 182 - 219 of T59832 P12, wherein said first, second, third and fourth amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of T59832 P 12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLV of T59832 P12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding far an edge portion of T59832 P12, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EC, having a structure as follows: a sequence starting from any of amino acid numbers 130-x to 130; and ending at any of amino acid numbers 131+ ((rr2) _ x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of T59832 P12, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence KPLEDQTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK in T59832 P12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P12, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYKTGNLYLRGPLKKSNA
PLVNVTLYYEALCGGCRAFLIRELFPTWLLVMEILNVTLVPYGNAQEQNVSGRWEFKC
QHGEEECKFNKVE corresponding to amino acids 1 - 130 of Q8WU77, which also corresponds to amino acids I - 130 of T59832 P12, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 162 - 250 of Q8WU77, which also corresponds to amino acids 131 - 219 of T59832 P12, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P 12, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise EC, having a structure as follows: a sequence starting from any of amino acid numbers 130-x to 130; and ending at any of amino acid numbers 131+ ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832_P18, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYK corresponding to amino acids 12 - 55 of GILT HUMAN, which also corresponds to amino acids 1 - 44 of T59832 P18, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 173 - 261 of GILT HUMAN, which also corresponds to amino acids 45 - 133 of T59832 P18, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P18, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise KC, having a structure as follows: a sequence starting from any of amino acid numbers 44-x to 44; and ending at any of amino acid numbers 45+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P18, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYK corresponding to amino acids 1 - 44 of Q8 WU77, which also corresponds to amino acids 1 - 44 of T59832 P I 8, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPWVTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 162 - 250 of Q8WU77, which also corresponds to amino acids 45 - 133 of T59832 P18, wherein said first and second amino acid sequences are contiguous and in a sequential order.
1 S According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P18, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise KC, having a structure as follows: a sequence starting from any of amino acid numbers 44-x to 44; and ending at any of amino acid numbers 45+ ((n-2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for T59832 P18, comprising a first amino acid sequence being at least 90 % homologous to MTLSPLLLFLPPLLLLLDVPTAAVQASPLQALDFFGNGPPVNYK corresponding to amino acids 1 - 44 of Q8NEI4, which also corresponds to amino acids 1 - 44 of T59832 P18, and a second amino acid sequence being at least 90 % homologous to CLQLYAPGLSPDTIMECAMGDRGMQLMHANAQRTDALQPPHEYVPW VTVNGKPLED
QTQLLTLVCQLYQGKKPDVCPSSTSSLRSVCFK corresponding to amino acids 162 - 250 of Q8NE14, which also corresponds to amino acids 45 - 133 of T59832 P18, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of T59832 P18, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about SO amino acids in length, wherein at least two amino acids comprise KC, having a structure as follows: a sequence starting from any of amino acid numbers 44-x to 44; and ending at any of amino acid numbers 45+ ((rr2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMGRPSE P4, comprising a first amino acid sequence being at least 90 % homologous to MRGSELPLVLLALVLCLAPRGRAVPLPAGGGTVLTKMYPRGNHWAVGHLMGKKSTG
ESSSVSERGSLKQQLREYIRWEEAARNLLGLIEAKENRNHQPPQPKALGNQQPSWDSED
SSNFKDVGSKGK corresponding to amino acids 1 - 127 of GRP-HUMAN, which also corresponds to amino acids 1 - 127 of HUMGRPSE P4, and a second amino acid sequence being at least 90 % homologous to GSQREGRNPQLNQQ corresponding to amino acids 148 of GRP-HUMAN, which also corresponds to amino acids 128 - 141 of HUMGRPSE
P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of HUMGRPSE P4, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise KG, having a structure as follows: a sequence starting from any of amino acid numbers 127-x to 127; and ending at any of amino acid numbers 128 + ((n-2) - x), in which x varies from 0 to n-2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HUMGRPSE P5, comprising a first amino acid sequence being at least 90 % homologous to MRGSELPLVLLALVLCLAPRGRAVPLPAGGGTVLTKMYPRGNHWAVGHLMGKKSTG
ESSSVSERGSLKQQLREYIRWEEAARNLLGLIEAKENRNHQPPQPKALGNQQPSWDSED
SSNFKDVGSKGK corresponding to amino acids 1 - I 27 of GRP_HUMAN, which also corresponds to amino acids I - 127 of HUMGRPSE P5, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DSLLQVLNVKEGTPS corresponding to amino acids 128 - 142 of HUMGRPSE P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of HUMGRPSE P5, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DSLLQVLNVKEGTPS in HUMGRPSE P5.
According to preferred embodiments of the present invention, there is provided an isolated chimerie polypeptide encoding for 811723 PEA_1 P6, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAGSPCRGLAPGREEQRALHKAGAVGGGVR
corresponding to amino acids 1 - 110 of 811723 PEA-1 P6, and a second amino acid sequence being at least 90 % homo logous to MYAQALLWGVLQRQAAAQHLHEHPPKLLRGHRVQERVDDRAEVEKRLREGEEDHV
RPEVGPRPVVLGFGRSHDPPNLVGHPAYGQCHNNQPWADTSRRERQRKEKHSMRTQ
corresponding to amino acids 1 - 112 of Q8IXM0, which also corresponds to amino acids 111 -222 of 811723 PEA_1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of Rl 1723 PEA_1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAGSPCRGLAPGREEQRALHKAGAVGGGVR of 811723 PEA 1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA_1 P6, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 1 - 83 of Q96AC2, which also corresponds to amino acids 1 - 83 of 811723 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ corresponding to amino acids 84 - 222 of 811723 PEA 1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLWGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPWLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ in 811723 PEA-1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA_1 P6, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 1 - 83 of Q8N2G4, which also corresponds to amino acids 1 - 83 of 811723 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGI-IRVQERVDDRAEVEKRLREGEEDI-IVRPEVGPRPVVLGFGRSI-IDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKI-ISMRTQ corresponding to amino acids 84 - 222 of 811723 PEA 1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ in 811723 PEA-1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA_1 P6, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 24 - 106 of BAC85518, which also corresponds to amino acids 1 - 83 of 811723 PEA_1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ corresponding to amino acids 84 - 222 of 811723 PEA_1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ in 811723 PEA-1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for Rl 1723 PEA_1 P7, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAG corresponding to amino acids 1 - 64 of Q96AC2, which also corresponds to amino acids 1 - 64 of 811723 PEA-1 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of R I 1723 PEA-1 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA_1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in 811723 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA_1 P7, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAG corresponding to amino acids 1 - 64 of Q8N2G4, which also corresponds to amino acids 1 - 64 ofRl 1723 PEA-1 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of 811723 PEA-1 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in 811723 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P7, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWVLG corresponding to amino acids 1 - 5 of 811723 PEA-1 P7, second amino acid sequence being at least 90 % homologous to IAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEVMEQSAG
corresponding to amino acids 22 - 80 of BAC85273, which also corresponds to amino acids 6 -64 of RI 1723 PEA-1 P7, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of 811723 PEA-1 P7, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of 811723 PEA-1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MWVLG of 811723 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA_1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in 811723 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA_1 P7, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAG corresponding to amino acids 24 - 87 of BAC85518, which also corresponds to amino acids 1 - 64 of 811723-PEA-1 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of 811723 PEA-1 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of Rl 1723 PEA-1 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in 811723 PEA 1 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P 13, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids I - 63 of Q96AC2, which also corresponds to amino acids 1 - 63 of 811723 PEA_1 P13, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DTKRTNTLLFEMRHFAKQLTT corresponding to amino acids 64 - 84 of 811723 PEA-1 P13, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P13, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DTKRTNTLLFEMRHFAKQLTT in 811723 PEA-1 P13.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P10, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids 1 - 63 of Q96AC2, which also corresponds to amino acids 1 - 63 of Rl 1723 PEA_1 P 10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of 811723 PEA_1 P10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in 811723 PEA 1 P10.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P10, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids 1 - 63 of Q8N2G4, which also corresponds to amino acids 1 - 63 of 811723 PEA-1 P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of 811723 PEA-1 P10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in 811723 PEA 1 P10.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P10, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWVLG corresponding to amino acids I - 5 of 811723 PEA-1 P10, second amino acid sequence being at least 90 % homologous to IAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVN VQDMCQKEVMEQSA
correspond ing to amino acids 22 - 79 of BAC85273, which also corresponds to amino acids 6 -63 of 811723 PEA-1 P 10, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of Rl 1723 PEA-1 P10, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a head of 811723 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence MWVLG of 811723 PEA 1 P10.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in 811723 PEA-1 PIO.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 811723 PEA-1 P10, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids 24 - 86 of BAC85518, which also corresponds to amino acids 1 - 63 of 811723 PEA-1 P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of R 1 1723 PEA-I-P 10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 811723 PEA-1 P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in 811723 PEA-1 P10.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for D56406 PEA-1 P2, comprising a first amino acid sequence being at least 90 % homologous to MMAGMKIQLVCMLLLAFSSWSLCSDSEEEMKALEADFLTNMHTSKISKAHVPSWKMT
LLNVCSLVNNLNSPAEETGEVHEEELVARRKLPTALDGFSLEAMLTIYQLHKICHSRAF
QHWE corresponding to amino acids 1 - 120 of NEUT HUMAN, which also corresponds to amino acids 1 - 120 of D56406 PEA-1 P2, second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence ARWLTPVIPALWEAETGGSRGQEMETIPANT corresponding to amino acids 121 - 151 of D56406 PEA-1 P2, and a third amino acid sequence being at least 90 %
homologous to LIQEDILDTGNDKNGKEEVIKRKIPYILKRQLYENKPRRPYILKRDSYYY corresponding to amino acids 121 - 170 of NEUT HUMAN, which also corresponds to amino acids 152 - 201 of D56406 PEA_1 P2, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of D56406_PEA_1 P2, comprising an amino acid sequence being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence encoding for ARWLTPVIPALWEAETGGSRGQEMETIPANT, corresponding to D56406 PEA 1 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for D56406 PEA-1 P5, comprising a first amino acid sequence being at least 90 % homologous to MMAGMKIQLVCMLLLAFSSWSLC
corresponding to amino acids 1 - 23 of NEUT_1-IUMAN, which also corresponds to amino acids 1 - 23 of D56406 PEA-1-P5, and a second amino acid sequence being at least 90 homologous to SEEEMKALEADFLTNMI-ITSKISKAHVPSWKMTLLNVCSLVNNLNSPAEETGEVHEEEL
VARRKLPTALDGFSLEAMLTIYQLHKICHSRAFQHWELIQEDILDTGNDKNGKEEVIKR
KIPYILKRQLYENKPRRPYILKRDSYYY corresponding to amino acid s 26 - 170 of NEUT HUMAN, which also corresponds to amino acids 24 - 168 of D56406 PEA-1 P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of D56406 PEA_1 P5, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise CS, having a I S structure as follows: a sequence starting from any of amino acid numbers 23-x to 24; and ending at any of amino acid numbers + ((rr2) - x), in which x varies from 0 to rr2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for D56406 PEA-I P6, comprising a first amino acid sequence being at least 90 % homologous to MMAGMKIQLVCMLLLAFSSWSLCSDSEEEMKALEADFLTNMHTSK corresponding to amino acids 1 - 45 of NEUT HUMAN, which also corresponds to amino acids 1 - 45 of D56406 PEA-1 P6, and a second amino acid sequence being at least 90 %
homologous to LIQEDILDTGNDKNGKEEVIKRKIPYILKRQLYENKPRRPYILKRDSYYY corresponding to amino acids 121 - 170 of NEUT_HUMAN, which also corresponds to amino acids 46 -95 of D56406 PEA 1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for an edge portion of D56406 PEA_1 P6, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise KL, having a structure as follows: a sequence starting from any of amino acid numbers 45-x to 46; and ending at any of amino acid numbers 46+ ((n-2) - x), in which x varies from 0 to r~2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H53393 PEA-1 P2, comprising a first amino acid sequence being at least 90 % homologous to MRTYRYFLLLFWVGQPYPTLSTPLSKRTSGFPAKKRALELSGNSKNELNRSKRSWMWN
QFFLLEEYTGSDYQYVGKLHSDQDRGDGSLKYILSGDGAGDLFIINENTGDIQATKRLD
QVTATDADDPTYGNSAKVVYSILQGQPYFSVESETGIIKTALLNMDRENREQYQWIQA
KDMGGQMGGLSGTTTVNITLTDVNDNPPRFPQSTYQFKTPESSPPGTPIGRIKASDADV
FLYLGPFKDSATVRIVVEDVDEPPVFSKLAYILQIREDAQINTTIGSVTAQDPDAARNPV
KYSVDRHTDMDRIFNIDSGNGSIFTSKLLDRETLLWHNITVIATEINNPKQSSRVPLYIKV
LDVNDNAPEFAEFYETFVCEKAKADQLIQTLHAVDKDDPYSGHQFSFSLAPEAASGSNF
TIQDNK corresponding to amino acids 1 - 543 of CAD6_HUMAN, which also corresponds to amino acids 1 - 543 of H53393 PEA-I P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GK
corresponding to amino acids 544 - 545 of H53393 PEA 1 P2, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H53393 PEA_1 P3, comprising a first amino acid sequence being at least 90 % homologous to MRTYRYFLLLFWGQPYPTLSTPLSKRTSGFPAKKRALELSGNSKNELNRSKRSWMWN
REEKPWILRAQAINRRTGRPVEPESEFIIKIHDINDNEPIFTKEVYTATVPEMSDVGTFW
QVTATDADDPTYGNSAKVVYSILQGQPYFSVESETGIIKTALLNMDRENREQYQWIQA
KDMGGQMGGLSGTTTVNITLTDVNDNPPRFPQSTYQFKTPESSPPGTPIGRIKASDADV
GENAEIEYSITDGEGLDMFDVITDQETQEGIITVKKLLDFEKKKVYTLKVEASNPWEPR
FLYLGPFKDSATVRIWEDVDEPPVFSKLAYILQIREDAQINTTIGSVTAQDPDAARNPV
LDVNDNAPEFAEFYETFVCEKAKADQ corresponding to amino acids 1 - 504 of CAD6 HUMAN, which also corresponds to amino acids 1 - 504 of H53393 PEA-1 P3, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence RFGFSLS corresponding to amino acids 505 - S 11 of H53393 PEA_I P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of H53393 PEA-1 P3, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence RFGFSLS
in H53393 PEA 1 P3.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for H53393 PEA-1 P6, comprising a first amino acid sequence being at least 90 % homologous to MRTYRYFLLLFWVGQPYPTLSTPLSKRTSGFPAKKRALELSGNSKNELNRSKRSWMWN
QFFLLEEYTGSDYQYVGKLHSDQDRGDGSLKYILSGDGAGDLFIINENTGDIQATKRLD
REEKPVYILRAQAINRRTGRPVEPESEFIIKIHDINDNEPIFTKEVYTATVPEMSDVGTFVV
QVTATDADDPTYGNSAKVVYSILQGQPYFSVESETGIIKTALLNMDRENREQYQWIQA
KDMGGQMGGLSGTTTVNITLTDVNDNPPRFPQSTYQFKTPESSPPGTPIGRIKASDADV
GENAEIEYSITDGEGLDMFDVITDQETQEGIITVKK corresponding to amino acids 1 - 333 of CAD6 HUMAN, which also corresponds to amino acids I - 333 of H53393 PEA-1 P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence VMPLLKHHTE corresponding to amino acids 334 -343 of H53393 PEA-1 P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of H53393 PEA-1 P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence VMPLLKHHTE in H53393 PEA 1 P6.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSU40434 PEA-1 P12, comprising a first amino acid sequence being at least 90 % homologous to MALPTARPLLGSCGTPALGSLLFLLFSLGWVQPSRTLAGETGQEAAPLDGVLANPPNISS
LSPRQLLGFPCAEVSGLSTERVRELAVALAQKNVKLSTEQLRCLAHRLSEPPEDLDALP
LDLLLFLNPDAFSGPQACTRFFSRITKANV DLLPRGAPERQRLLPAALACWGVRGSLLS
EADVRALGGLACDLPGRFVAESAEVLLPRLVSCPGPLDQDQQEAARAALQGGGPPYGP
PSTWSVSTMDALRGLLPVLGQPIIRSIPQGIVAAWRQRSSRDPSWRQPERTILRPRFRRE
VEKTACPSGKKAREIDESLIFYKKWELEACVDAALLATQMDRVNAIPFTYEQLDVLKH
KLDELYPQGYPESVIQHLGYLFLKMSPEDIRKWNVTSLETLKALLEVNKGHEMSPQVA
TLIDRFVKGRGQLDKDTLDTLTAFYPGYLCSLSPEELSSVPPSSIW corresponding to amino acids 1 - 458 of Q14859, which also corresponds to amino acids 1 - 458 of HSU40434 PEA 1 P 12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for HSU40434 PEA-1 P12, comprising a first amino acid sequence being at least 90 % homologous to MALPTARPLLGSCGTPALGSLLFLLFSLGWVQPSRTLAGETGQ corresponding to amino acids I - 43 of Q9BTR2, which also corresponds to amino acids I - 43 of HSU40434 PEA-1 P 12, second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence E corresponding to amino acids 44 - 44 of HSU40434 PEA-1 P12, and a third amino acid sequence being at least 90 %
homologous to AAPLDGVLANPPNISSLSPRQLLGFPCAEVSGLSTERVRELAVALAQKNVKLSTEQLRC
LAHRLSEPPEDLDALPLDLLLFLNPDAFSGPQACTRFFSRITKANVDLLPRGAPERQRLL
PAALACWGVRGSLLSEADVRALGGLACDLPGRFVAESAEVLLPRLVSCPGPLDQDQQE
AARAALQGGGPPYGPPSTWSVSTMDALRGLLPVLGQPIIRSIPQGIVAAWRQRSSRDPS
WRQPERTILRPRFRREVEKTACPSGKKAREIDESLIFYKKWELEACVDAALLATQMDRV
NAIPFTYEQLDVLKHKLDELYPQGYPESVIQHLGYLFLKMSPEDIRKWNVTSLETLKAL
LEVNKGHEMSPQVATLIDRFVKGRGQLDKDTLDTLTAFYPGYLCSLSPEELSSVPPSSIW
corresponding to amino acids 44 - 457 of Q9BTR2, which also corresponds to amino acids 45 -458 ofHSU40434 PEA-1 P12, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for an edge portion of HSU40434 PEA_ 1 P 12, comprising an amino acid sequence being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at.least about 90% and most preferably at least about 95%
homologous to the sequence encoding for E, corresponding to HSU40434 PEA 1 P12.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P2, comprising a first amino acid sequence being at least 90 % homologous to MLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNRTFIWD
VKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATVVRIGTFCSNGTVSRIKMQ
EGVKMALHLPWFHPRNVSGFSIANRSSIKRLCIIESVFEGEGSATLMSANYPEGFPEDEL
MTWQFVVPAHLRASVSFLNFNLSNCERKEERVEYYIPGSTTNPEVFKLEDKQPGNMAG
NFNLSLQGCDQDAQSPGILRLQFQVLVQHPQNES corresponding to amino acids 67 - 341 of Q8WU91, which also corresponds to amino acids 1 - 275 of M77904 P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NKIYVVDLSNERAMSLTIEPRPVKQSRKFVPGCFVC LESRTCSSNLTLTSGSKHKISFLCD
DLTRLWMNVEKTISCTDHRYCQRKSYSLQVPSDILHLPVELHDFSWKLLVPKDRLSLVL
VPAQKLQQHTHEKPCNTSFSYLVASAIPSQDLYFGSFCPGGSIKQIQVKQNISVTLRTFAP
SFQQEASRQGLTVSFIPYFKEEGVFTVTPDTKSKVYLRTPNWDRGLPSLTSVSWNISVPR
DQVACLTFFKERSGWCQTGRAFMIIQEQRTRAEEIFSLDEDVLPKPSFHHHSFWVNISN
CSPTSGKQLDLLFSVTLTPRTVDLTVILIAAVGGGVLLLSALGLIICCVKKKKKKTNKGP
AVGIYNGNINTEMPRQPKKFQKGRKDNDSHVYAVIEDTMWGHLLQDSSGSFLQPEVD
TYRPFQGTMGVCPPSPPTICSRAPTAKLATEEPPPRSPPESESEPYTFSHPNNGDVSSKDT
DIPLLNTQEPMEPAE corresponding to amino acids 276 - 770 of M77904 P2, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NKIYVVDLSNERAMSLTIEPRPVKQSRKFVPGCFVCLESRTCSSNLTLTSGSKHKISFLCD
VPAQKLQQHTHEKPCNTSFSYLVASAIPSQDLYFGSFCPGGSIKQIQVKQNISVTLRTFAP
SFQQEASRQGLTVSFIPYFKEEGVFTVTPDTKSKVYLRTPNWDRGLPSLTSVSWNISVPR
DQVACLTFFKERSGVVCQTGRAFMIIQEQRTRAEEIFSLDEDVLPKPSFHHHSFWVNISN
CSPTSGKQLDLLFSVTLTPRTVDLTVILIAAVGGGVLLLSALGLIICCVKKKKKKTNKGP
AVGIYNGNINTEMPRQPKKFQKGRKDNDSHVYAVIEDTMVYGHLLQDSSGSFLQPEVD
TYRPFQGTMGVCPPSPPTICSRAPTAKLATEEPPPRSPPESESEPYTFSHPNNGDVSSKDT
DIPLLNTQEPMEPAE in M77904 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P2, comprising a first amino acid sequence being at least 90 % homologous to MLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNRTFIWD
VKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATV VRIGTFCSNGTVSRIKMQ
EGVKMALHLPWFHPRNVSGFSIANRSSIKRLCIIESVFEGEGSATLMSANYPEGFPEDEL
MTWQFVVPAHLRASVSFLNFNLSNCERKEERVEYYIPGSTTNPEVFKLEDKQPGNMAG
NFNLSLQGCDQDAQSPGILRLQFQVLVQHPQNESNKIYVVDLSNERAMSLTIEPRPVKQ
SRKFVPGCFVCLESRTCSSNLTLTSGSKHKISFLCDDLTRLWMNVEKTISCTDHRYCQR
KSYSLQVPSDILHLPVELHDFSWKLLVPKDRLSLVLVPAQKLQQHTHEKPCNTSFSYLV
ASAIPSQDLYFGSFCPGGSIKQIQVKQNISVTLRTFAPSFQQEASRQGLTVSFIPYFKEEGV
FTVTPDTKSKVYLRTPNWDRGLPSLTSVSWNISVPRDQVACLTFFKERSGVVCQTGRAF
MIIQEQRTRAEEIFSLDEDVLPKPSFHHHSFWVNISNCSPTSGKQLDLLFSVTLTPRTVDL
TVILIAAVGGGVLLLSALGLIICCVKKKKKKTNKGPAVGIYNGNINTEMPRQPKKFQKG
RKDNDSHVYAVIEDTMVYGHLLQDSSGSFLQPEVDTYRPFQGTMGVCPPSPPTICSRAP
TAKLATEEPPPRSPPESESEPYTFSHPNNGDVSSKDTDIPLLNTQEPMEPAE corresponding to amino acids 67 - 836 of Q96QU7, which also corresponds to amino acids 1 -770 of M77904 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P4, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGVSIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
KRHITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATV VRIGTFCSNGTVSR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKRLCIIESVFEGEGSATLMSANYPEGFP
EDELMTWQFVVPAHLRASVSFLNFNLSNCERKEERVEYYIPGSTTNPEVFKLEDKQPGN
MAGNFNLSLQGCDQDAQSPGILRLQFQVLVQHPQNES corresponding to amino acids 1 -341 of Q8WU91, which also corresponds to amino acids 1 - 341 of M77904 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence NKIYWDLSNERAMSLTIEPRPVKQSRKFVPGCFVCLESRTCSSNLTLTSGSKHKISFLCD
DLTRLWMNVEKTISTPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLT
PVIPALWEAKAGGSLEVRSSRPAWPTW corresponding to amino acids 342 - 487 of M77904 P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail ofM77904 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence NKIYWDLSNERAMSLTIEPRPVKQSRKFVPGCFVC LESRTCSSNLTLTSGSKHKISFLCD
DLTRLWMNVEKTISTPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLT
PVIPALWEAKAGGSLEVRSSRPAWPTW in M77904 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P4, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGVSIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
KRHITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATWRIGTFCSNGTVSR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKRLCIIESVFEGEGSATLMSANYPEGFP
EDELMTWQFVVPAHLRASVSFLNFNLSNCERKEERVEYYIPGSTTNPEVFKLEDKQPGN
MAGNFNLSLQGCDQDAQSPGILRLQFQVLVQHPQNESNKIYVVDLSNERAMSLTIEPRP
VKQSRKFVPGCFVCLESRTCSSNLTLTSGSKHKISFLCDDLTRLWMNVEKTIS
corresponding to amino acids I - 416 of Q9HSV8, which also corresponds to amino acids 1 -416 of M77904 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence TPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLTPVIPALWEAKAGGS
LEVRSSRPAWPTW corresponding to amino acids 417 - 487 of M77904 P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about I S 90% and most preferably at least about 95% homologous to the sequence TPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLTPVIPALWEAKAGGS
LEVRSSRPAWPTW in M77904 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P4, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGVSIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
KRI-IITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATVVRIGTFCSNGTVSR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKRLCIIESVFEGEGSATLMSANYPEGFP
EDELMTWQFWPAHLRASVSFLNFNLSNCERKEERVEYYIPGSTTNPEVFKLEDKQPGN
MAGNFNLSLQGCDQDAQSPGILRLQFQVLVQHPQNESNKIYVVDLSNERAMSLTIEPRP
VKQSRKFVPGCFVCLESRTCSSNLTLTSGSKHKISFLCDDLTRLWMNVEKTIS
corresponding to amino acids 1 - 416 of Q96QU7, which also corresponds to amino acids 1 -416 of M77904 P4, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95%
homologous to a polypeptide having the sequence TPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLTPVIPALWEAKAGGS
LEVRSSRPAWPTW corresponding to amino acids 417 - 487 of M77904 P4, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P4, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence TPLNQCICPWPWIALLSPPCLSGVPWVGCKSYQKGPSGRARWLTPVIPALWEAKAGGS
LEVRSSRPAWPTW in M77904 P4.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P5, comprising a first amino acid sequence being at least 90 % homologous to MIIQEQRTRAEEIFSLDEDVLPKPSFHHHSFWVNISNCSPTSGKQLDLLFSVTLTPRTVDL
TVILIAAVGGGVLLLSALGLIICCVKKKKKKTNKGPAVGIYNGNINTEMPRQPKKFQKG
RKDNDSHVYAVIEDTMVYGHLLQDSSGSFL,QPEVDTYRPFQGTMGVCPPSPPTICSRAP
TAKLATEEPPPRSPPESESEPYTFSHPNNGDVSSKDTDIPLLNTQEPMEPAE corresponding to amino acids 606 - 836 of Q96QU7, which also corresponds to amino acids 1 -231 of M77904 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P5, comprising a first amino acid sequence being at least 90 % homologous to MIIQEQRTRAEEIFSLDEDVLPKPSFHHHSFWVNISNCSPTSGKQLDLLFSVTLTPRTVDL
TVILIAAVGGGVLLLSALGLIICCVKKKKKKTNKGPAVGIYNGNINTEMPRQPKKFQKG
RKDNDSHVYAVIEDTMVYGHLLQDSSGSFLQPEVDTYRPFQGTMGVCPPSPPTICSRAP
TAKLATEEPPPRSPPESESEPYTFSHPNNGDVSSKDTDIPLLNTQEPMEPAE corresponding to amino acids 419 - 649 of Q9H8C2, which also corresponds to amino acids 1 -231 of M77904 P5.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P7, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGVSIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
18o KRHITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATVVRIGTFCSNGTVSR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKR corresponding to amino acids 1 - 219 of Q8WU91, which also corresponds to amino acids 1 - 219 ofM77904 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence EKAPPCYLIRLKHTRSSLF corresponding to amino acids 220 - 238 of M77904 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence EKAPPCYLIRLKHTRSSLF in M77904 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P7, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGVSIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
KRHITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATVVRIGTFCSNGTVSR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKR corresponding to amino acids 1 - 219 of Q9HSV8, which also corresponds to amino acids 1 - 219 of M77904 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence EKAPPCYLIRLKHTRSSLF corresponding to amino acids 220 - 238 of M77904 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence EKAPPCYLIRLKHTRSSLF in M77904 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for M77904 P7, comprising a first amino acid sequence being at least 90 % homologous to MAGLNCGV SIALLGVLLLGAARLPRGAEAFEIALPRESNITVLIKLGTPTLLAKPCYIVIS
KRHITMLSIKSGERIVFTFSCQSPENHFVIEIQKNIDCMSGPCPFGEVQLQPSTSLLPTLNR
TFIWDVKAHKSIGLELQFSIPRLRQIGPGESCPDGVTHSISGRIDATV VRIGTFCSNGTV SR
IKMQEGVKMALHLPWFHPRNVSGFSIANRSSIKR corresponding to amino acids 1 - 219 of Q96QU7, which also corresponds to amino acids 1 - 219 of M77904 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence EKAPPCYLIRLKHTRSSLF corresponding to amino acids 220 - 238 of M77904 P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of M77904 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence EKAPPCYLIRLKHTRSSLF in M77904 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 225299 PEA 2 P2, comprising a first amino acid sequence being at least 90 % homologous to MKSSGLFPFLVLLALGTLAPWAVEGSGKSFKAGVCPPKKSAQCLRYKKPECQSDWQCP
GKKRCCPDTCGIKCLDPVDTPNPTRRKPGKCPVTYGQCLMLNPPNFCEMDGQCKRDLK
CCMGMCGKSCVSPVK corresponding to amino acids 1 - 131 of ALK1 HUMAN, which also corresponds to amino acids 1 - 131 of 225299 PEA 2 P2, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GKQGMRAH corresponding to amino acids 132 - 139 of 225299 PEA 2 P2, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 225299 PEA 2 P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homobgous to the sequence GKQGMRAH in 225299 PEA 2 P2.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 225299 PEA 2 P3, comprising a first amino acid sequence being at least 90 % homologous to MKSSGLFPFLVLLALGTLAPWAVEGSGKSFKAGVCPPKKSAQCLRYKKPECQSDWQCP
GKKRCCPDTCGIKCLDPVDTPNPTRRKPGKCPVTYGQCLMLNPPNFCEMDGQCKRDLK
CCMGMCGKSCVSPVK corresponding to amino acids 1 - 131 of ALKI HUMAN, which also corresponds to amino acids 1 - 131 of 225299 PEA 2 P3, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence GEKRHHKQLRDQEVDPLEMRRHSAG corresponding to amino acids 132 - 156 of 225299 PEA 2 P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 225299 PEA 2 P3, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence GEKRHHKQLRDQEVDPLEMRRHSAG in 225299 PEA 2 P3.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 225299 PEA 2 P7, comprising a first amino acid sequence being at least 90 % homologous to MKSSGLFPFLVLLALGTLAPWAVEGSGKSFKAGVCPPKKSAQCLRYKKPECQSDWQCP
GKKRCCPDTCGIKCLDPVDTPNP corresponding to amino acids 1 - 81 of ALKI HUMAN, which also corresponds to amino acids I - 81 of 225299 PEA 2 P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence RGSLGSAQ corresponding to amino acids 82 - 89 of 225299 PEA 2-P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
According to preferred embodiments of the present invention, there is provided an isolated polypeptide encoding for a tail of 225299 PEA 2 P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence RGSLGSAQ in 225299 PEA 2 P7.
According to preferred embodiments of the present invention, there is provided an isolated chimeric polypeptide encoding for 225299 PEA 2 PIO, comprising a first amino acid sequence being at least 90 % homologous to MKSSGLFPFLVLLALGTLAPWAVEGSGKSFKAGVCPPKKSAQCLRYKKPECQSDWQCP
GKKRCCPDTCGIKCLDPVDTPNPT corresponding to amino acids 1 - 82 ofALKI HUMAN, which also corresponds to amino acids 1 - 82 of 225299 PEA 2 P10.
According to preferred embodiments of the present invention, there is provided an antibody capable of specifically binding to an epitope of an amino acid sequence as described herein.
Optionally the amino acid sequence corresponds to a bridge, edge portion, tail, head or insertion as described herein.
Optionally the antibody is capable of differentiating between a splice variant having said epitope and a corresponding known protein.
According to preferred embodiments of the present invention, there is provided a kit for detecting ovarian cancer, comprising a kit detecting overexpression of a splice variant as described herein.
Optionally the kit comprises a NAT-based technology.
Optionally the kit further comprises at least one primer pair capable of selectively hybridizing to a nucleic acid sequence as described herein.
Optionally the kit further comprises at least one oligonucleotide capable of selectively hybridizing to a nucleic acid sequence as described herein.
Optionally the kit comprises an antibody as described herein.
Optionally the kit further comprises at least one reagent for performing an ELISA or a Western blot.
According to preferred embodiments of the present invention, there is provided a method for detecting ovarian cancer, comprising detecting overexpression of a splice variant as described herein.
Optionally detecting overexpression is performed with a NAT-based technology.
Optionally detecting overexpression is performed with an immunoassay.
Optionally the irmnunoassay comprises an antibody as described herein.
According to preferred embodiments of the present invention, there is provided a biomaiker capable of detecting ovarian cancer, comprising any of the above nucleic acid sequences or a fragment thereof, or any of the above amino acid sequences or a fragment thereof.
According to preferred embodiments of the present invention, there is provided a method for screening for ovarian cancer, comprising detecting ovarian cane er cells with a biomarker or an antibody or a method or assay as described herein.
According to preferred embodiments of the present invention, there is provided a method for diagnosing ovarian cancer, comprising detecting ovarian cancer cells with a biomarker or an antibody or a method or assay as described herein.
According to preferred embodiments of the present invention, there is provided a method for monitoring disease progression and/or treatment efficacy and/or relapse of ovarian cancer, comprising detecting ovarian cancer cells with a biomarker or an antibody or a method or assay as described herein.
According to preferred embodiments of the present invention, there is provided a method of selecting a therapy for ovarian cancer, comprising detecting ovarian cancer cells with a biomarker or an antibody or a method or assay as described herein and selecting a therapy according to said detection.
According to preferred embodiments of the present invention, preferably any of the above nucleic acid and/or amino acid sequences further comprises any sequence having at least about 70%, preferably at least about 80%, more preferably at least about 90%, most preferably at least about 95% homology thereto.
Unless otherwise noted, all experimental data relates to variants of the present invention, named according to the segment being tested (as expression was tested through RT-PCR as described).
All nucleic acid sequences and/or amino acid sequences shown herein as embodiments of the present invention relate to their isolated form, as isolated polynucleotides (including for all transcripts), oligonucleotides (including for all segments, amplicons and primers), peptides (including for all tails, bridges, insertions or heads, optionally including other antibody epitopes as described herein) and/or polypeptides (including for all proteins). It should be noted that oligonucleotide and polynucleotide, or peptide and polypeptide, may optionally be used interchangeably.
BRIEF DESCRIPTION OF DRAWINGS
Figure 1 is schematic summary of cancer biomarkers selection engine and the wet validation stages.
Figure 2. Schematic illustration, depicting grouping of transcripts of a given cluster based on presence or absence of unique sequence regions.
Figure 3 is schematic summary of quantitative real-time PCR analysis.
Figure 4 is schematic presentation of the oligonucleotide based microarray fabrication.
Figure 5 is schematic summary of the oligonucleotide based microarray experimental flow.
Figure 6 shows cancer and cell-line vs. normal tissue expression for .
Figure 7 shows expression of segment8 in H61775 in cancerous vs. norrcancerous tissues.
Figure 8 shows expression of segment8 in H61775 in normal tissues.
Figure 9 shows cancer and cell-line vs. normal tissue expression.
Figure 10 is a histogram showing over expression of T juncl 1-17 transcripts in cancerous ovary samples relative to the normal samples.
Figure 11 is a histogram showing expression of T juncl 1-17 transcripts in normal tissues.
Figure 12 shows cancer and cell-line vs. normal tissue expression.
Figure 13 is a histogram showing over expression of HUMGRPSEjunc3-7 transcripts in cancerous ovary samples relative to the normal samples.
Figure 14 is a histogram showing expression of HUMGRPSEjunc3-7 transcripts in normal tissues.
Figure 15 shows cancer and cell-line vs. normal tissue expression.
Figure 16 is a histogram showing over expression of 811723 segl3 transcripts in S cancerous ovary samples relative to the normal PM samples.
Figure 17 is a histogram showing expression of RI 1723 segl3 transcripts in normal tissue samples.
Figure 18 is a histogram showing over expression of R 11723 junc 11-18 transcripts in cancerous ovary samples relative to the normal samples.
Figure 19 is a histogram showing expression of 811723 juncl l-18 transcripts in normal tissue samples.
Figure 20 shows cancer and cell-line vs. normal tissue expression.
Figure 21 is a histogram showing over expression of H53393 segl3 transcripts in cancerous ovary samples relative to the normal samples.
I S Figure 22 is a histogram showing over expression of H53393 junc2l-22 transcripts in cancerous ovary samples relative to the normal samples.
Figure 23 shows cancer and cell-line vs. normal tissue expression.
Figure 24 shows cancer and cell-line vs. normal tissue expression.
Figure 25 shows cancer and cell-line vs. normal tissue expression.
Figure 26 is a histogram showing over expression of 225299 juncl3-14-21 transcripts in cancerous ovary samples relative to the normal samples.
Figures 27A and 27B are histograms showing over expression of 225299 seg20 transcripts in cancerous ovary samples relative to the normal samples (27A) or in normal tissues (27B).
Figures 28A and 28B are histograms showing over expression of 225299 seg23 transcripts in cancerous ovary samples relative to the normal samples (28A) or in normal tissues (28B).
Figure 29 shows cancer and cell-line vs. normal tissue expression.
Figure 30 is a histogram showing down regulation of T39971 june23-338 transcripts in cancerous ovary samples relative to the normal samples.
Figure 31 is a histogram showing expression of T39971 junc23-33R transcripts in normal tissues.
Figure 32 shows cancer and cell-line vs. normal tissue expression.
Figures 33A and 33B are histograms showing down regulation of 244808 junc8-11 transcripts in cancerous ovary samples relative to the normal samples (33A) or expression in normal tissues (33B).
Figure 34 shows cancer and cell-line vs. normal tissue expression.
Figure 35 shows cancer and cell-line vs. normal tissue expression.
Figure 36 shows cancer and cell-line vs. normal tissue expression.
Figure 37 shows cancer and cell-line vs. normal tissue expression.
Figure 38 shows cancer and cell-line vs. normal tissue expression.
Figure 39 shows cancer and cell-line vs. normal tissue expression.
Figure 40 shows cancer and cell-line vs. normal tissue expression.
Figure 41 shows cancer and cell-line vs. normal tissue expression.
Figure 42 shows cancer and cell-line vs. normal tissue expression.
Figure 43 is a histogram showing differential expression of a variety of transcripts in cancerous ovary samples relative to the normal samples.
Figure 44 shows cancer and cell-line vs. normal tissue expression.
DESCRIPTION OF PREFERRED EMBODIMENTS
The present invention is of novel markers for ovarian cancer that are both sensitive and accurate. Biomolecular sequences (amino acid and/or nucleic acid sequences) uncovered using the methodology of the present invention and described herein can be efficiently utilized as tissue or pathological markers and/or as drugs or drug targets for treating or preventing a disease.
Furthermore, at least certain of these markers are able to distinguish between various types of ovarian cancer, such as Ovarian epithelial tumors (serous, mucinous, endometroid, clear cell, and Brenner tumor), ovarian germ-cell tumors, (teratoma, dysgerminoma, endodermal sinus tumor, and embryonal carcinoma) and ovarian stromal tumors (originating from granulosa, theca, Sertoli, Leydig, and collagen-producing stromal cells), alone or in combination. These markers are differentially expressed, and preferably overexpressed in ovarian cancer specifically, as opposed to normal ovarian tissue. The measurement of these markers, alone or in combination, in patient samples provides information that the diagnostician can correlate with a probable diagnosis of ovarian cancer. The markers of the present invention, alone or in combination, show a high degree of differential detection between ovarian cancer and non-cancerous states.
The markers of the present invention, alone or in combination, can be used for prognosis, prediction, screening, early diagnosis, staging, therapy selection and treatment monitoring of ovarian cancer. For example, optionally and preferably, these markers may be used for staging ovarian cancer and/or monitoring the progression of the disease. Furthermore, the markers of the present invention, alone or in combination, can be used for detection of the source of metastasis found in anatomical places other thenovary. Also, one or more of the markers may optionally be used in combination with one or more other ovarian cancer markers (other than those described herein). According to an optional embodiment of the present invention, such a combination may be used to differentiate between various types of ovarian cancer, such as Ovarian epithelial tumors (serous, mucinous, endometroid, clear cell, and Brenner tumor), ovarian gerrrrcell tumors, (teratoma, dysgerminoma, endodermal sinus tumor, and embryonal carcinoma) and ovarian stromal tumors (originating from either granulosa, theca, Sertoli, Leydig, and collagerrproducing stromal cells).
These markers are specifically released to the bloodstream under conditions of ovarian cancer (or one of the above indicative conditions), and/or are otherwise expressed at a much higher level and/or specifically expressed in ovarian cancer tissue or cells, and/or tissue or cells under one of the above indicative conditions. The measurement of these markers, alone or in combination, in patient samples provides information that the diagnostician can correlate with a probable diagnosis of ovarian cancer and/or a condition that it is indicative of a higher risk for ovarian cancer.
The present invention therefore also relates to diagnostic assays for ovarian cancer, and methods of use of such markers for detection of ovarian cancer, optionally and preferably in a sample taken from a subject (patient), which is more preferably some type of blood sample.
In another embodiment, the present invention relates to bridges, tails, heads and/or insertions, and/or analogs, homologs and derivatives of such peptides. Such bridges, tails, heads and/or insertions are described in greater detail below with regard to the Examples.
As used herein a "tail" refers to a peptide sequence at the end of an amino acid sequence that is unique to a splice variant according to the present invention.
Therefore, a splice variant having such a tail may optionally be considered as a chimera, in that at least a first portion ofthe splice variant is typically highly homologous (often 100% identical) to a portion of the corresponding known protein, while at least a second portion of the variant comprises the tail.
As used herein a "head" refers to a peptide sequence at the beginning of an amino acid sequence that is unique to a splice variant according to the present invention. Therefore, a splice variant having such a head may optionally be considered as a chimera, in that at least a first portion of the splice variant comprises the head, while at least a second portion is typically highly homologous (often 100% identical) to a portion of the corresponding known protein.
As used herein "an edge portion" refers to a connection between two portions of a splice variant according to the present invention that were not joined in the wild type or known protein. An edge may optionally arise due to a join between the above "known protein" portion of a variant and the tail, for example, and/or may occur if an internal portion of the wild type sequence is no longer present, such that two portions of the sequence are now contiguous in the splice variant that were not contiguous in the known protein. A "bridge" may optionally be an edge portion as described above, but may also include a join between a head and a "known protein" portion of a variant, or a join between a tail and a "known protein"
portion of a variant, or a join between an insertion and a "known protein" portion of a variant.
Optionally and preferably, a bridge between a tail or a head or a unique insertion, and a "known protein" portion of a variant, comprises at least about 10 amino acids, more preferably at least about 20 amino acids, most preferably at least about 30 amino acids, and even more preferably at least about 40 amino acids, in which at least one amino acid is from the tail/head/insertion and at least one amino acid is from the "known protein"
portion of a variant.
Also optionally, the bridge may comprise any number of amino acids from about 10 to about 40 amino acids (for example, 10, 11, 12, 13...37, 38, 39, 40 amino acids in length, or any number in between).
It should be noted that a bridge cannot be extended beyond the length of the sequence in either direction, and it should be assumed that every bridge description is to be read in such manner that the bridge length does not extend beyond the sequence itself.
Furthermore, bridges are described with regard to a sliding window in certain contexts below. For example, certain descriptions of the bridges feature the following format: a bridge between two edges (in which a portion of the known protein is not present in the variant) may optionally be described as follows: a bridge portion of CONTIG-NAME PI
(representing the name of the protein), comprising a polypeptide having a length "n", wherein n is at least about amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most 10 preferably at least about 50 amino acids in length, wherein at least two amino acids comprise XX (2 amino acids in the center of the bridge, one from each end of the edge), having a structure as follows (numbering according to the sequence of CONTIG-NAME P I
): a sequence starting from any of amino acid numbers 49-x to 49 (for example); and ending at any of amino acid numbers 50 + ((n-2) - x) (for example), in which x varies from 0 to n-2.
In this example, it I 5 should also be read as including bridges in which n is any number of amino acids between 10-50 amino acids in length. Furthermore, the bridge polypeptide cannot extend beyond the sequence, so it should be read such that 49-x (for example) is not less than 1, nor 50 +
((r~2) - x) (for example) greater than the total sequence length.
In another embodiment, this invention provides antibodies specifically recognizing the splice variants and polypeptide fragments thereof of this invention.
Preferably such antibodies differentially recognize splice variants of the present invention but do not recognize a corresponding known protein (such known proteins are discussed with regard to their splice variants in the Examples below).
In another embodiment, this invention provides an isolated nucleic acid molecule encoding for a splice variant according to the present invention, having a nucleotide sequence as set forth in any one of the sequences listed herein, or a sequence complementary thereto. In another embodiment, this invention provides an isolated nucleic acid molecule, having a nucleotide sequence as set forth in any one of the sequences listed herein, or a sequence complementary thereto. In another embodiment, this invention provides an oligonucleotide of at least about 12 nucleotides, specifically hybridizable with the nucleic acid molecules of this invention. In another embodiment, this invention provides vectors, cells, liposomes and compositions comprising the isolated nucleic acids of this invention.
In another embodiment, this invention provides a method for detecting a splice variant according to the present invention in a biological sample, comprising:
contacting a biological sample with an antibody specifically recognizing a splice variant according to the present invention under conditions whereby the antibody specifically interacts with the splice variant in the biological sample but do not recognize known corresponding proteins (wherein the known protein is discussed with regard to its splice variants) in the Examples below), and detecting said interaction; wherein the presence of an interaction correlates with the presence of a splice variant in the biological sample.
In another embodiment, this invention provides a method for detecting a splice variant nucleic acid sequences in a biological sample, comprising: hybridizing the isolated nucleic acid molecules or oligonucleotide fragments of at least about a minimum length to a nucleic acid material of a biological sample and detecting a hybridization complex; wherein the presence of a hybridization complex correlates with the presence of a splice variant nucleic acid sequence in the biological sample.
According to the present invention, the splice variants described herein are norrlimiting examples of markers for diagnosing ovarian cancer. Each splice variant marker of the present invention can be used alone or in combination, for various uses, including but not limited to, prognosis, prediction, screening, early diagnosis, determination of progression, therapy selection and treatment monitoring of ovarian cancer.
According to optional but preferred embodiments of the present invention, any marker according to the present invention may optionally be used alone or combination. Such a combination may optionally comprise a plurality of markers described herein, optionally including any subcombination of markers, and/or a combination featuring at least one other marker, for example a known marker. Furthermore, such a combination may optionally and preferably be used as described above with regard to determining a ratio between a quantitative or semi-quantitative measurement of any marker described herein to any other marker described herein, and/or any other known marker, and/or any other marker. With regard to such a ratio between any marker described herein (or a combination thereof) and a known marker, more preferably the known marker comprises the "known protein" as described in greater detail below with regard to each cluster or gene.
According to other preferred embodiments of the present invention, a splice variant protein or a fragment thereof, or a splice variant nucleic acid sequence or a fragment thereof, may be featured as a biomarker for detecting ovarian cancer and/or an indicative condition, such that a biomarker may optionally comprise any of the above.
According to still other preferred embodiments, the present invention optionally and preferably encompasses any amino acid sequence or fragment thereof encoded by a nucleic acid sequence corresponding to a splice variant protein as described herein. Any oligopeptide or peptide relating to such an amino acid sequence or fragment thereof may optionally also (additionally or alternatively) be used as a biomarker, including but not limited to the unique amino acid sequences of these proteins that are depicted as tails, heads, insertions, edges or bridges. The present invention also optionally encompasses antibodies capable of recognizing, and/or being elicited by, such oligopeptides or peptides.
The present invention also optionally and preferably encompasses any nucleic acid sequence or fragment thereof, or amino acid sequence or fragment thereof, corresponding to a splice variant of the present invention as described above, optionally for any application.
Non-limiting examples of methods or assays are described below.
The present invention also relates to kits based upon such diagnostic methods or assays.
Nucleic acid sequences and Oligonucleotides Various embodiments of the present invention encompass nucleic acid sequences described hereinabove; fragments thereof, sequences hybridizable therewith, sequences homologous thereto, sequences encoding similar polypeptides with different codon usage, altered sequences characterized by mutations, such as deletion, insertion or substitution of one or more nucleotides, either naturally occurring or artificially induced, either randomly or in a targeted fashion.
The present invention encompasses nucleic acid sequences described herein;
fragments thereof, sequences hybridizable therewith, sequences homologous thereto [e.g., at least 50 %, at least 55 %, at least 60%, at least 65 %, at least 70 %, at least 75 %, at least 80 %, at least 85 %, at least 95 % or more say 100 % identical to the nucleic acid sequences set forth below], sequences encoding similar polypeptides with different codon usage, altered sequences characterized by mutations, such as deletion, insertion or substitution of one or more nucleotides, either naturally occurring or man induced, either randomly or in a targeted fashion. The present invention also encompasses homologous nucleic acid sequences (i.e., which form a part of a polynucleotide sequence of the present invention) which include sequence regions unique to the polynucleotides of the present invention.
In cases where the polynucleotide sequences of the present invention encode previously unidentified polypeptides, the present invention also encompasses novel polypeptides or portions thereof, which are encoded by the isolated polynucleotide and respective nucleic acid fragments thereof described hereinabove.
A "nucleic acid fragment" or an "oligonucleotide" or a "polynucleotide" are used herein interchangeably to refer to a polymer of nucleic acids. A polynucleotide sequence of the present invention refers to a single or double stranded nucleic acid sequences which is isolated and provided in the form of an RNA sequence, a complementary polynucleotide sequence (cDNA), a genomic polynucleotide sequence and/or a composite polynucleotide sequences (e.g., a combination of the above).
As used herein the phrase "complementary polynucleotide sequence" refers to a sequence, which results from reverse transcription of messenger RNA using a reverse transcriptase or any other RNA dependent DNA polymerase. Such a sequence can be subsequently amplified in vivo or in vitro using a DNA dependent DNA
polymerase.
As used herein the phrase "genomic polynucleotide sequence" refers to a sequence derived (isolated) from a chromosome and thus it represents a contiguous portion of a chromosome.
As used herein the phrase "composite polynucleotide sequence" refers to a sequence, which is composed of genomic and cDNA sequences. A composite sequence can include some exonal sequences required to encode the polypeptide of the present invention, as well as some intronic sequences interposing therebetween. The intronic sequences can be of any source, including of other genes, and typically will include conserved splicing signal sequences. Such intronic sequences may further include cis acting expression regulatory elements.
Preferred embodiments of the present invention encompass oligonucleotide probes.
An example of an oligonucleotide probe which can be utilized by the present invention is a single stranded polynucleotide which includes a sequence complementary to the unique sequence region of any variant according to the present invention, including but not limited to a nucleotide sequence coding for an amino sequence of a bridge, tail, head and/or insertion according to the present invention, and/or the equivalent portions of any nucleotide sequence given herein (including but not limited to a nucleotide sequence of a node, segment or amplicon described herein).
Alternatively, an oligonucleotide probe of the present invention can be designed to hybridize with a nucleic acid sequence encompassed by any of the above nucleic acid sequences, particularly the portions specified above, including but not limited to a nucleotide sequence coding for an amino sequence of a bridge, tail, head and/or insertion according to the present invention, and/or the equivalent portions of any nucleotide sequence given herein (including but not limited to a nucleotide sequence of a node, segment or amplicon described herein).
Oligonucleotides designed according to the teachings of the present invention can be generated according to any oligonucleotide synthesis method known in the art such as enzymatic synthesis or solid phase synthesis. Equipment and reagents for executing solid-phase synthesis are commercially available from, for example, Applied Biosystems. Any other means for such synthesis may also be employed; the actual synthesis of the oligonucleotides is well within the capabilities of one skilled in the art and can be accomplished via established methodologies as detailed in, for example, "Molecular Cloning: A laboratory Manual" Sambrook et al., (1989);
"Current Protocols in Molecular Biology" Volumes I-III Ausubel, R. M., ed.
(1994); Ausubel et al., "Current Protocols in Molecular Biology", John Wiley and Sons, Baltimore, Maryland (1989); Perbal, "A Practical Guide to Molecular Cloning", John Wiley & Sons, New York (1988) and "Oligonucleotide Synthesis" Gait, M. J., ed. (1984) utilizing solid phase chemistry, e.g. cyanoethyl phosphoramidite followed by deprotection, desalting and purification by for example, an automated trityl-on method or HPLC.
Oligonucleotides used according to this aspect of the present invention are those having a length selected from a range of about 10 to about 200 bases preferably about 15 to about 150 bases, more preferably about 20 to about 100 bases, most preferably about 20 to about 50 bases.
Preferably, the oligonucleotide of the present invention features at least 17, at least 18, at least 19, at least 20, at least 22, at least 25, at least 30 or at least 40, bases specifically hybridizable with the biomarkers of the present invention.
The oligonucleotides of the present invention may comprise heterocylic nucleosides consisting ofpurines and the pyrimidines bases, bonded in a 3' to 5' phosphodiester linkage.
Preferably used oligonucleotides are those modified at one or more of the backbone, internucleoside linkages or bases, as is broadly described hereinunder.
Specific examples of preferred oligonucleotides useful according to this aspect of the present invention include oligonucleotides containing modified backbones or norrnatural internucleoside linkages. Oligonucleotides having modified backbones include those that retain a phosphorus atom iri the backbone, as disclosed in U.S. Pat. NOs: 4,469,863;
4,476,301;
5,023,243; 5,177,196; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717;
5,321,131;
5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466, 677; 5,476,925; 5,519,126;
5,536,821;
5,541,306; 5,550,111; 5,563,253; 5,571,799; 5,587,361; and 5,625,050.
Preferred modified oligonucleotide backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkyl phosphotriesters, methyl and other alkyl phosphonates including 3'-alkylene phosplnnates and chiral phosphonates, phosphinates, phosphoramidates including 3'-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates having normal 3'-5' linkages, 2'-5' linked analogs of these, and those having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'. Various salts, mixed salts and free acid forms can also be used.
Alternatively, modified oligonucleotide backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatom and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside);
siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones;
methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S and CHI component parts, as disclosed in U.S. Pat. Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134;
5,216,141; 5,235,033;
5,264,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; 5,489,677;
5,541,307;
5,561,225; 5,596,086; 5,602,240; 5,610,289; 5,602,240; 5,608,046; 5,610,289;
5,618,704; 5,623, 070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439.
Other oligonucleotides which can be used according to the present invention, are those modified in both sugar and the internucleoside linkage, i.e., the backbone, of the nucleotide units are replaced with novel groups. The base units are maintained for complementation with the appropriate polynucleotide target. An example for such an oligonucleotide mimetic, includes peptide nucleic acid (PNA). United States patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, each of which is herein incorporated by reference. Other backbone modifications, which can be used in the present invention are disclosed in U.S. Pat. No: 6,303,374.
Oligonucleotides of the present invention may also include base modifications or substitutions. As used herein, "unmodified" or "natural" bases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U).
Modified bases include but are not limited to other synthetic and natural bases such as S-methylcytosine (5-me-C), S~hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly S-bromo, ~trifluoromethyl and other S-substituted uracils and cytosines, '~methylguanine and '~methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine.
Further bases particularly useful for increasing the binding affinity of the oligomeric compounds of the invention include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, including 2 aminopropyladenine, S~propynyluracil and ~propynylcytosine.
5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6 1.2 °C and are presently preferred base substitutions, even more particularly when combined with 2'-O-methoxyethyl sugar modifications.
Another modification of the oligonucleotides of the invention involves chemically linking to the oligonucleotide one or more moieties or conjugates, which enhance the activity, cellular distribution or cellular uptake of the oligonucleotide. Such moieties include but are not limited to lipid moieties such as a cholesterol moiety, cholic acid, a thioether, e.g., hexyl-S-tritylthiol, a thiocholesterol, an aliphatic chain, e.g., dodecandiol or undecyl residues, a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate, a polyamine or a polyethylene glycol chain, or adamantine acetic acid, a palmityl moiety, or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety, as disclosed in U.S. Pat. No: 6,303,374.
It is not necessary for all positions in a given oligonucleotide molecule to be uniformly modified, and in fact more than one of the aforement Toned modifications may be incorporated in a single compound or even at a single nucleoside within an oligonucleotide.
It will be appreciated that oligonucleotides of the present invention may include further modifications for more efficient use as diagnostic agents and/or to increase bioavailability, therapeutic efficacy and reduce cytotoxicity.
To enable cellular expression of the polynucleotides of the present invention, a nucleic acid construct according to the present invention may be used, which includes at least a coding region of one of the above nucleic acid sequences, and further includes at least one cis acting regulatory element. As used herein, the phrase "cis acting regulatory element"
refers to a polynucleotide sequence, preferably a promoter, which binds a trans acting regulator and regulates the transcription of a coding sequence located downstream thereto.
Any suitable promoter sequence can be used by the nucleic acid construct of the present invention.
Preferably, the promoter utilized by the nucleic acid construct of the present invention is active in the specific cell population transformed. Examples of cell type-specific and/or tissue-specific promoters include promoters such as albumin that is liver specific, lymphoid specific promoters [Calame et al., (1988) Adv. Immunol. 43:235-275]; in particular promoters of T cell receptors [Winoto et al., (1989) EMBO J. 8:729-733] and immunoglobulins;
[Banerji et al.
(1983) Cell 33729-740], neurorrspeciflc promoters such as the neurofilament promoter [Byrne et al. (1989) Proc. Natl. Acid. Sci. USA 86:5473-5477], pancreas-specific promoters [Edlunch et al. (1985) Science 230:912-916] or mammary gland-specific promoters such as the milk whey promoter (U.S. Pat. No. 4,873,316 and European Application Publication No. 264,166).
The nucleic acid construct of the present invention can further include an enhancer, which can be adjacent or distant to the promoter sequence and can function in up regulating the transcription therefrom.
The nucleic acid construct of the present invention preferably further includes an appropriate selectable marker and/or an origin of replication. Preferably, the nucleic acid construct utilized is a shuttle vector, which can propagate both in E. coli (wherein the construct comprises an appropriate selectable marker and origin of replication) and be compatible for propagation in cells, or integration in a gene and a tissue of choice. The construct according to the present invention can be, for example, a plasmid, a bacmid, a phagemid, a cosmid, a phage, a virus or an artificial chromosome.
Examples of suitable constructs include, but are not limited to, pcDNA3, pcDNA3.1 (+/-), pGL3, PzeoSV2 (+/-), pDisplay, pEF/myc/cyto, pCMV/myc/cyto each of which is commercially available from Invitrogen Co. (www.invitrogen.com). Examples of retroviral I S vector and packaging systems are those sold by Clontech, San Diego, Calif., includingRetro-X
vectors pLNCX and pLXSN, which permit cloning into multiple cloning sites and the trasgene is transcribed from CMV promoter. Vectors derived from Mo-MuLV are also included such as pBabe, where the transgene will be transcribed from the 5'LTR promoter.
Currently preferred in vivo nucleic acid transfer techniques include transfection with viral or norrviral constructs, such as adenovirus, lentivirus, Herpes simplex I virus, or adeno associated virus (AAV) and lipid-based systems. Useful lipids for lipid-mediated transfer of the gene are, for example, DOTMA, DOPE, and DC-Chol [Tonkinson et al., Cancer Investigation, 14(1): 54-65 (1996)]. The most preferred constructs for use in gene therapy are viruses, most preferably adenoviruses, AAV, lentiviruses, or retroviruses. A viral construct such as a retroviral construct includes at least one transcriptional promoter/enhancer or locus-defining element(s), or other elements that control gene expression by other means such as alternate splicing, nuclear RNA export, or post-translational modification of messenger.
Such vector constructs also include a packaging signal, long terminal repeats (LTRs) or portions thereof, and positive and negative strand primer binding sites appropriate to the virus used, unless it is already present in the viral construct. In addition, such a construct typically includes a signal sequence for secretion of the peptide from a host cell in which it is placed.
Preferably the signal sequence for this purpose is a mammalian signal sequence or the signal sequence of the polypeptide variants of the present invention. Optionally, the construct may also include a signal that directs polyadenylation, as well as one or more restriction sites and a translation termination sequence. By way of example, such constructs will typically include a 5' LTR, a tRNA binding site, a packaging signal, an origin of second-strand DNA
synthesis, and a 3' LTR
or a portion thereof. Other vectors can be used that are norrviral, such as cationic lipids, polylysine, and dendrimers.
Hybridization assays Detection of a nucleic acid of interest in a biological sample may optionally be effected by hybridizatior~based assays using an oligonucleotide probe (non-limiting examples of probes according to the present invention were previously described).
Traditional hybridization assays include PCR, RT-PCR, Real-time PCR, RNase protection, in-situ hybridization, primer extension, Southern blots (DNA
detection), dot or slot blots (DNA, RNA), and Northern blots (RNA detection) (NAT type assays are described in greater detail below). More recently, PNAs have been described (Nielsen et al.
1999, Current Opin. Biotechnol. 10:71-75). Other detection methods include kits containing probes on a dipstick setup and the like.
Hybridization based assays which allow the detection of a variant of interest (i.e., DNA
or RNA) in a biobgical sample rely on the use of oligonucleotides which can be 10, 15, 20, or to 100 nucleotides long preferably from 10 to 50, more preferably from 40 to 50 nucleotides long.
Thus, the isolated polynucleotides (oligonucleotides) of the present invention are preferably hybridizable with any of the herein described nucleic acid sequences under moderate 25 to stringent hybridization conditions.
Moderate to stringent hybridization conditions are characterized by a hybridization solution such as containing 10 % dextrane sulfate, 1 M NaCI, 1 % SDS and 5 x 106 cpm 32P
labeled probe, at 65 °C, with a final wash solution of 0.2 x SSC and 0.1 % SDS and final wash at 65°C and whereas moderate hybridization is effected using a hybridization solution 30 containing 10 % dextrane sulfate, 1 M NaCI, 1 % SDS and 5 x 106 cpm 32P
labeled probe, at 65 °C, with a final wash solution of 1 x SSC and 0.1 % SDS and final wash at SO °C.
More generally, hybridization of short nucleic acids (below 200 by in length, e.g. 17-40 by in length) can be effected using the following exemplary hybridization protocols which can be modified according to the desired stringency; (i) hybridization solution of 6 x SSC and 1 SDS or 3 M TMACI, 0.01 M sodium phosphate (pH 6.8), 1 mM EDTA (pH 7.6), 0.5 %
SDS, 100 pg/ml denatured salmon sperm DNA and 0.1 % nonfat dried milk, hybridization temperature of 1 - 1.5 °C below the TIn, final wash solution of 3 M TMACI, 0.01 M
sodium phosphate (pH
6.8), 1 mM EDTA (pH 7.6), 0.5 % SDS at 1 - 1.5 °C below the Tm; (ii) hybridization solution of 6 x SSC and 0.1 % SDS or 3 M TMACI, 0.01 M sodium phosphate (pl-I 6.8), 1 mM EDTA
(pH 7.6), 0.5 % SDS, 100 pg/ml denatured salmon sperm DNA and 0.1 % nonfat dried milk, hybridization temperature of 2 - 2.5 °C below the Tm, final wash solution of 3 M TMACI, 0.01 M sodium phosphate (pH 6.8), 1 mM EDTA (pH 7.6), 0.5 % SDS at 1 - 1.5 °C below the Tm, final wash solution of 6 x SSC, and final wash at 22 °C; (iii) hybridization solution of 6 x SSC
and 1 % SDS or 3 M TMACI, 0.01 M sodium phosphate (pH 6.8), 1 mM EDTA (pH
7.6), 0.5 SDS, 100 pg/ml denatured salmon sperm DNA and 0.1 % nonfat dried milk, hybridization temperature.
The detection of hybrid duplexes can be carried out by a number of methods.
Typically, hybridization duplexes are separated from unhybridized nucleic acids and the labels bound to the duplexes are then detected. Such labels refer to radioactive, fluorescent, biological or enzymatic tags or labels of standard use in the art. A label can be conjugated to either the oligonucleotide probes or the nucleic acids derived from the biological sample.
Probes can be labeled according to numerous well known methods. Non-limiting examples of radioactive labels include 3H, 14C, 32P, and 35S. Norrlimiting examples of detectable markers include ligands, fluorophores, chemiluminescent agents, enzymes, and antibodies. Other detectable markers for use with probes, which can enable an increase in sensitivity of the method of the invention, include biotin and radio-nucleotides. It will become evident to the person of ordinary skill that the choice of a particular label dictates the manner in which it is bound to the probe.
For example, oligonucleotides of the present invention can be labeled subsequent to synthesis, by incorporating biotinylated dNTPs or rNTP, or some similar means (e.g., photo cross-linking a psoralen derivative of biotin to RNAs), followed by addition of labeled streptavidin (e.g., phycoerythrin-conjugated streptavidin) or the equivalent.
Alternatively, when fluorescently-labeled oligonucleotide probes are used, fluorescein, lissamine, phycoerythrin, rhodamine (Perkin Elmer Cetus), Cy2, Cy3, Cy3.5, CyS, Cy5.5, Cy7, FIuorX
(Amersham) and others [e.g., Kricka et al. (1992), Academic Press San Diego, CalifJ can be attached to the oligonucleotides.
Those skilled in the art will appreciate that wash steps may be employed to wash away excess target DNA or probe as well as unbound conjugate. Further, standard heterogeneous assay formats are suitable for detecting the hybrids using the labels present on the oligonucleotide primers and probes.
It will be appreciated that a variety of controls may be usefully employed to improve accuracy of hybridization assays. For instance, samples may be hybridized to an irrelevant probe and treated with RNAse A prior to hybridization, to assess false hybridization.
Although the present invention is not specifically dependent on the use of a label for the detection of a particular nucleic acid sequence, such a label might be beneficial, by increasing the sensitivity of the detection. Furthermore, it enables automation. Probes can be labeled according to numerous well known methods.
As commonly known, radioactive nucleotides can be incorporated into probes of the invention by several methods. Norr limiting examples of radioactive labels include 3H, '4C, 32P, and 35S.
Those skilled in the art will appreciate that wash steps may be employed to wash away excess target DNA or probe as well as unbound conjugate. Further, standard hetero~neous assay formats are suitable for detecting the hybrids using the labels present on the oligonucleotide primers and probes.
It will be appreciated that a variety of controls may be usefully employed to improve accuracy of hybridization assays.
Probes of the invention can be utilized with naturally occurnng sugar-phosphate backbones as well as modified backbones including phosphorothioates, dithionates, alkyl phosphonates and a-nucleotides and the like. Probes of the invention can be constructed of either ribonucleic acid (RNA) or deoxyribonucleic acid (DNA), and preferably of DNA.
NAT Assays Detection of a nucleic acid of interest in a biological sample may also optionally be effected by NAT-based assays, which involve nucleic acid amplification ~chnology, such as PCR for example (or variations thereof such as real-time PCR for example).
As used herein, a "primer" defines an oligonucleotide which is capable of annealing to (hybridizing with) a target sequence, thereby creating a double stranded region which can serve as an initiation point for DNA synthesis under suitable conditions.
Amplification of a selected, or target, nucleic acid sequence may be carried out by a number of suitable methods. See generally Kwoh et al., 1990, Am. Biotechnol.
Lab. 8:14 Numerous amplification techniques have been described and can be readily adapted to suit particular needs of a person of ordinary skill. Norr limiting examples of amplification techniques include polymerase chain reaction (PCR), ligase chain reaction (LCR), strand displacement amplification (SDA), transcriptiorrbased amplification, the q3 replicase system and NASBA
(Kwoh et al., 1989, Proc. NatI. Acad. Sci. USA 86, 1173-1177; Lizardi et al., 1988, BioTechnology 6:1197-1202; Malek et al., 1994, Methods Mol. Biol., 28:253-260;
and Sambrook et al., 1989, supra).
The terminology "amplification pair" (or "primer pair") refers herein to a pair of oligonucleotides (oligos) of the present invention, which are selected to be used together in amplifying a selected nucleic acid sequence by one of a number of types of amplification processes, preferably a polymerase chain reaction. Other types of amplification processes include ligase chain reaction, strand displacement amplification, or nucleic acid sequence-based amplification, as explained in greater detail below. As commonly known in the art, the oligos are designed to bind to a complementary sequence under selected conditions.
In one particular embodiment, amplification of a nucleic acid sample from a patient is amplified under conditions which favor the amplification of the most abundant differentially expressed nucleic acid. In one preferred embodiment, RT PCR is carried out on an mRNA
sample from a patient under conditions which favor the amplification of the most abundant mRNA. In another preferred embodiment, the amplification of the differentially expressed nucleic acids is carried out simultaneously. It will be realized by a person skilled in the art that such methods could be adapted for the detection of differentially expressed proteins instead of differentially expressed nucleic acid sequences.
The nucleic acid (i.e. DNA or RNA) for practicing the present invention may be obtained according to well known methods.
Oligonucleotide primers of the present invention may be of any suitable length, depending on the particular assay format and the particular needs and targeted genomes employed. Optionally, the oligonucleotide primers are at least 12 nucleotides in length, preferably between 15 and 24 molecules, and they may be adapted to be especially suited to a chosen nucleic acid amplification system. As commonly known in the art, the oligonucleotide primers can be designed by taking into consideration the melting point of hybridization thereof with its targeted sequence (Sambrook et al., 1989, Molecular Cloning -A
Laboratory Manual, 2nd Edition, CSH Laboratories; Ausubel et al., 1989, in Current Protocols in Molecular Biology, John Wiley & Sons Inc., N.Y.).
It will be appreciated that antisense oligonucleotides may be employed to quantify expression of a splice isoform of interest. Such detection is effected at the pre-mRNA level.
Essentially the ability to quantitate transcription from a splice site of interest can be effected based on splice site accessibility. Oligonucleotides may compete with splicing factors for the splice site sequences. Thus, low activity of the antisense oligonucleotide is indicative of splicing activity.
The polymerise chain reaction and other nucleic acid amplification reactions are well known in the art (various norr limiting examples of these reactions are described in greater detail below). The pair of oligonucleotides according to this aspect of the present invention are preferably selected to have compatible melting temperatures (Tm), e.g., melting temperatures which differ by less than that 7 °C, preferably less than 5 °C, more preferably less than 4 °C, most preferably less than 3 °C, ideally between 3 °C and 0 °C.
Polymerise Chain Reaction (PCR): The polymerise chain reaction (PCR), as described in U.S. Pat. Nos. 4,683,195 and 4,683,202 to Mullis and Mullis et al., is a method of increasing the concentration of a segment of target sequence in a mixture of genomic DNA
without cloning or purification. This technology provides one approach to the problems of low target sequence concentration. PCR can be used to directly increase the concentration of the target to an easily detectable level. This process for amplifying the target sequence involves the introduction of a molar excess of two oligonucleotide primers which are complementary to their respective strands of the double-stranded target sequence to the DNA mixture containing the desired target sequence. The mixture is denatured and then allowed to hybridize. Following hybridization, the primers are extended with polymerise so as to form complementary strands. The steps of denaturation, hybridization (annealing), and polymerise extension (elongation) can be repeated as often as needed, in order to obtain relatively high concentrations of a segment of the desired target sequence.
The length of the segment of the desired target sequence is determined by the relative positions of the primers with respect to each other, and, therefore, this length is a controllable parameter. Because the desired segments of the target sequence become the dominant sequences (in terms of concentration) in the mixture, they are said to be "PCR-amplified."
Ligase Chain Reaction (LCR or LAR): The ligase chain reaction [LCR; sometimes referred to as "Ligase Amplification Reaction" (LAR)] has developed into a well-recognized alternative method of amplifying nucleic acids. In LCR, four oligonucleotides, two adjacent oligonucleotides which uniquely hybridize to one strand of target DNA, and a complementary set of adjacent oligonucleotides, which hybridize to the opposite strand are mixed and DNA ligase is added to the mixture. Provided that there is complete complementarity at the junction, ligase will covalently link each set of hybridized molecules. Importantly, in LCR, two probes are ligated together only when they base-pair with sequences in the target sample, without gaps or mismatches. Repeated cycles of denaturation, and ligation amplify a short segment of DNA.
LCR has also been used in combination with PCR to achieve enhanced detection of single-base changes: see for example Segev, PCT Publication No. W09001069 A1 (1990).
However, because the four oligonucleotides used in this assay can pair to form two short ligatable fragments, there is the potential for the generation of target independent background signal. The use of LCR for mutant screening is limited to the examination of specific nucleic acid positions.
Self-Sustained Synthetic Reaction (3SRlNASBA): The self sustained sequence replication reaction (3SR) is a transeriptiorrbased in vitro amplification system that can exponentially amplify RNA sequences at a uniform temperature. The amplified RNA can then be utilized for mutation detection. In this method, an oligonucleotide primer is used to add a phage RNA
polymerise promoter to the 5' end of the sequence of interest. In a cocktail of enzymes and substrates that includes a second primer, reverse transcriptase, RNase H, RNA
polymerise and ribo-and deoxyribonucleoside triphosphates, the target sequence undergoes repeated rounds of transcription, cDNA synthesis and second-strand synthesis to amplify the area of interest. The use of 3SR to detect mutations is kinetically limited to screening small segments of DNA (e.g., 200-300 base pairs).
Q-Beta (Q(3) Replicase: In this method, a probe which recognizes the sequence of interest is attached to the replicatable RNA template for Q[3 replicase. A
previously identified major problem with false positives resulting from the replication of unhybridized probes has been addressed through use of a sequence-specific ligation step. However, available thermostable DNA ligases are not effective on this RNA substrate, so the ligation must be performed by T4 DNA ligase at low temperatures (37 degrees C.). This prevents the use of high temperature as a means of achieving specificity as in the LCR, the ligation event can be used to detect a mutation at the junction site, but not elsewhere.
A successful diagnostic method must be very specific. A straight-forward method of controlling the specificity of nucleic acid hybridization is by controlling the temperature of the reaction. While the 3SR/NASBA, and Q(3 systems are all able to generate a large quantity of signal, one or more of the enzymes involved in each cannot be used at high temperature (i.e., >
55 degrees C). Therefore the reaction temperatures cannot be raised to prevent non-specific hybridization of the probes. If probes are shortened in order to make them melt more easily at low temperatures, the likelihood of having more than one perfect match in a complex genome increases. For these reasons, PCR and LCR currently dominate the research field in detection technologies.
The basis of the amplification procedure in the PCR and LCR is the fact that the products of one cycle become usable templates in all subsequent cycles, consequently doubling the population with each cycle. The final yield of any such doubling system can be expressed as:
(1+X)n ~, where "X" is the mean efficiency (percent copied in each cycle), "n"
is the number of cycles, and "y" is the overall efficiency, or yield of the reaction. If every copy of a target DNA is utilized as a template in every cycle of a polymerase chain reaction, then the mean efficiency is 100 %. If 20 cycles of PCR are performed, then the yield will be 220, or 1,048,576 copies of the starting material. If the reaction conditions reduce the mean efficiency to 85 %, then the yield in those 20 cycles will be only 1.8520, or 220,513 copies of the starting material. In other words, a PCR running at 85 % efficiency will yield only 21 % as much final product, compared to a reaction running at 100 % efficiency. A reaction that is reduced to 50 % mean efficiency will yield less than 1 % of the possible product.
In practice, routine polymerase chain reactions rarely achieve the theoretical maximum yield, and PCRs are usually run for more than 20 cycles to compensate for the lower yield. At 50 % mean efficiency, it would take 34 cycles to achieve the milliorrfold amplification theoretically possible in 20, and at lower efficiencies, the number of cycles required becomes prohibitive. In addition, any background products that amplify with a better mean efficiency than the intended target will become the dominant products.
Also, many variables can influence the mean efficiency of PCR, including target DNA
length and secondary structure, primer length and design, primer and dNTP
concentrations, and buffer composition, to name but a few. Contamination of the reaction with exogenous DNA
(e.g., DNA spilled onto lab surfaces) or cross-contamination is also a major consideration.
Reaction conditions must be carefully optimized for each different primer pair and target sequence, and the process can take days, even for an experienced investigator.
The laboriousness of this process, including numerous technical considerations and other factors, presents a significant drawback to using PCR in the clinical setting. Indeed, PCR has yet to penetrate the clinical market in a significant way. The same concerns arise with LCR, as LCR
must also be optimized to use different oligonucleotide sequences for each target sequence. In addition, both methods require expensive equipment, capable of precise temperature cycling.
Many applications of nucleic acid detection technologies, such as in studies of allelic variation, involve not only detection of a specific sequence in a complex background, but also the discrimination between sequences with few, or single, nucleotide differences. One method of the detection of allele-specific variants by PCR is based upon the fact that it is difficult for Taq polymerase to synthesize a DNA strand when there is a mismatch between the template strand and the 3' end of the primer. An allele-specific variant may be detected by the use of a primer that is perfectly matched with only one of the possible alleles; the mismatch to the other allele acts to prevent the extension of the primer, thereby preventing the amplification of that sequence.
This method has a substantial limitation in that the base composition of the mismatch influences the ability to prevent extension across the mismatch, and certain mismatches do not prevent extension or have only a minimal effect.
A similar 3'-mismatch strategy is used with greater effect to prevent ligation in the LCR.
Any mismatch effectively blocks the action of the thermostable ligase, but LCR
still has the drawback of target-independent background ligation products initiating the amplification.
Moreover, the combination of PCR with subsequent LCR to identify the nucleotides at individual positions is also a clearly cumbersome proposition for the clinical laboratory.
The direct detection method according to various preferred embodiments of the present invention may be, for example a cycling probe reaction (CPR) or a branched DNA
analysis.
When a sufficient amount of a nucleic acid to be detected is available, there are advantages to detecting that sequence directly, instead of making more copies of that target, (e.g., as in PCR and LCR). Most notably, a method that does not amplify the signal exponentially is more amenable to quantitative analysis. Even if the signal is enhanced by attaching multiple dyes to a single oligonucleotide, fie correlation between the final signal intensity and amount of target is direct. Such a system has an additional advantage that the products of the reaction will not themselves promote further reaction, so contamination of lab 1 S surfaces by the products is not as much of a concern. Recently devised techniques have sought to eliminate the use of radioactivity and/or improve the sensitivity in automatable formats. Two examples are the "Cycling Probe Reaction" (CPR), and "Branched DNA" (bDNA).
Cycling probe reaction (CPR): The cycling probe reaction (CPR), uses a long chimeric oligonucleotide in which a central portion is made of RNA while the two termini are made of DNA. Hybridization of the probe to a target DNA and exposure to a thermostable RNase H
causes the RNA portion to be digested. 'This destabilizes the remaining DNA
portions of the duplex, releasing the remainder of the probe from the target DNA and allowing another probe molecule to repeat the process. The signal, in the form of cleaved probe molecules, accumulates at a linear rate. While the repeating process increases the signal, the RNA
portion of the oligonucleotide is vulnerable to RNases that may carried through sample preparation.
Branched DNA: Branched DNA (bDNA), involves oligonucleotides with branched structures that allow each individual oligonucleotide to carry 35 to 40 labels (e.g., alkaline phosphatase enzymes). While this enhances the signal from a hybridization event, signal from nonspecific binding is similarly increased.
The detection of at least one sequence -change according to various preferred embodiments of the present invention may be accomplished by, for example restriction fragment length polymorphism (RFLP analysis), allele specific oligonucleotide (ASO) analysis, Denaturing/Temperature Gradient Gel Electrophoresis (DGGE/TGGE), Single-Strand Conformation Polymorphism (SSCP) analysis or Dideoxy fingerprinting (ddF).
The demand for tests which allow the detection of specific nucleic acid sequences and sequence changes is growing rapidly in clinical diagnostics. As nucleic acid sequence data for genes from humans and pathogenic organisms accumulates, the demand for fast, cost-effective, and easy to-use tests for as yet mutations within specific sequences is rapidly increasing.
A handful of methods have been devised to scan nucleic acid segments for mutations.
One option is to determine the entire gene sequence of each test sample (e.g., a bacterial isolate).
For sequences under approximately 600 nucleotides, this may be accomplished using amplified material (e.g., PCR reaction products). This avoids the time and expense associated with cloning the segment of interest. However, specialized equipment and highly trained personnel are required, and the method is too labor-intense and expensive to be practical and effective in the clinical setting.
In view of the difficulties associated with sequencing, a given segment of nucleic acid may be characterized on several other levels. At the lowest resolution, the size of the molecule can be determined by electrophoresis by comparison to a known standard run on the same gel. A
more detailed picture of the molecule may be achieved by cleavage with combinations of restriction enzymes prior to electrophoresis, to allow construction of an ordered map. The presence of specific sequences within the fragment can be detected by hybridization of a labeled probe, or the precise nucleotide sequence can be determined by partial chemical degradation or by primer extension in the presence of chain-terminating nucleotide analogs.
Restriction fragment length polymorphism (RFLP): For detection of single-base differences between like sequences, the requirements of the analysis are often at the highest level of resolution. For cases in which the position of the nucleotide in question is known in advance, several methods have been developed for examining single base changes without direct sequencing. For example, if a mutation of interest happens to fall within a restriction recognition sequence, a change in the pattern of digestion can be used as a diagnostic tool (e.g., restriction fragment length polymorphism [RFLP] analysis).
Single point mutations have been also detected by the creation or destruction of RFLPs.
Mutations are detected and bcalized by the presence and size of the RNA
fragments generated by cleavage at the mismatches. Single nucleotide mismatches in DNA
heteroduplexes are also recognized and cleaved by some chemicals, providing an alternative strategy to detect single base substitutions, generically named the "Mismatch Chemical Cleavage" (MCC).
However, this method requires the use of osmium tetroxide and piperidine, two highly noxious chemicals which are not suited for use in a clinical laboratory.
RFLP analysis suffers from low sensitivity and requires a large amount of sample. When RFLP analysis is used for the detection of point mutations, it is, by its nature, limited to the detection of only those single base changes which fall within a restriction sequence of a known restriction endonuclease. Moreover, the majority of the available enzymes have 4 to 6 base-pair recognition sequences, and cleave too frequently for many large-scale DNA
manipulations.
Thus, it is applicable only in a small fraction of cases, as most mutations do not fall within such sites.
A handful of rare-cutting restriction enzymes with 8 base-pair specificities have been isolated and these are widely used in genetic mapping, but these enzymes are few in number, are I S limited to the recognition of G+C-rich sequences, and cleave at sites that tend to be highly clustered. Recently, endonucleases encoded by group I introns have been discovered that might have greater than 12 base-pair specificity, but again, these are few in number.
Allele specific oligonucleotide (ASO): If the change is not in a recognition sequence, then allele-specific oligonucleotides (ASOs), can be designed to hybridize in proximity to the mutated nucleotide, such that a primer extension or ligation event can bused as the indicator of a match or a mis-match. Hybridization with radioactively labeled allelic specific oligonucleotides (ASO) also has been applied to the detection of specific point mutations. The method is based on the differences in the melting temperature of short DNA fragments differing by a single nucleotide. Stringent hybridization and washing conditions can differentiate between mutant and wild-type alleles. The ASO approach applied to PCR products also has been extensively utilized by various researchers to detect and characterize point mutations in ras genes and gsp/gip oncogenes. Because of the presence of various nucleotide changes in multiple positions, the ASO method requires the use of many oligonucleotides to cover all possible oncogenic mutations.
With either of the techniques described above (i.e., RFLP and ASO), the precise location of the suspected mutation must be known in advance of the test. That is to say, they are inapplicable when one needs to detect the presence of a mutation within a gene or sequence of interest.
DenaturinglTemperature Gradient Gel Electrophoresis (DGGElTGGE): Two other methods rely on detecting changes in electrophoretic mobility in response to minor sequence changes. One of these methods, termed "Denaturing Gradient Gel Electrophoresis" (DGGE) is based on the observation that slightly different sequences will display different patterns of local melting when electrophoretically resolved on a gradient gel. In this manner, variants can be distinguished, as differences in melting properties of homoduplexes versus heteroduplexes differing in a single nucleotide can detect the presence of mutations in the target sequences because of the corresponding changes in their electrophoretic mobilities. The fragments to be analyzed, usually PCR products, are "clamped" at one end by a long stretch of CtC base pairs (30-80) to allow complete denaturation of the sequence of interest without complete dissociation of the strands. The attachment of a GC "clamp" to the DNA fragments increases the faction of mutations that can be recognized by DGGE. Attaching a GC clamp to one primer is critical to ensure that the amplified sequence has a low dissociation temperature.
Modifications of the technique have been developed, using temperature gradients, and the method can be also applied to RNA:RNA duplexes.
Limitations on the utility of DGGE include the requirement that the denaturing conditions must be optimized for each type of DNA to be tested. Furthermore, the method requires specialized equipment b prepare the gels and maintain the needed high temperatures during electrophoresis. The expense associated with the synthesis of the clamping tail on one oligonucleotide for each sequence to be tested is also a major consideration.
In addition, long running times are required for DGGE. The long running time of DGGE was shortened in a modification of DGGE called constant denaturant gel electrophoresis (CDGE).
CDGE requires that gels be performed under different denaturant conditions in order to reach high efficiency for the detection of mutations.
A technique analogous to DGGE, termed temperature gradient gel electrophoresis (TGGE), uses a thermal gradient rather than a chemical denaturant gradient.
TGGE requires the use of specialized equipment which can generate a temperature gradient perpendicularly oriented relative to the electrical field. TGGE can detect mutations in relatively small fragments of DNA
therefore scanning of large gene segments requires the use of multiple PCR
products prior to running the gel.
Single-Strand Conformation Polymorphism (SSCP): Another common method, called "Single-Strand Conformation Polymorphism" (SSCP) was developed by Hayashi, Sekya and colleagues and is based on the observation that single strands of nucleic acid can take on characteristic conformations in non-denaturing conditions, and these conformations influence electrophoretic mobility. The complementary strands assume sufficiently different structures that one strand may be resolved from the other. Changes in sequences within the fragment will also change the conformation, consequently altering the mobility and allowing this to be used as an assay for sequence variations.
The SSCP process involves denaturing a DNA segment (e.g., a PCR product) that is labeled on both strands, followed by slow electrophoretic separation on a norrdenaturing polyacrylamide gel, so that intra-molecular interactions can form and not be disturbed during the run. This technique is extremely sensitive to variations in gel composition and temperature. A
serious limitation of this method is the relative difficulty encountered in comparing data generated in different laboratories, under apparently similar conditions.
Dideoxy fingerprinting (ddF): The dideoxy fingerprinting (ddF) is another technique developed to scan genes for the presence of mutations. The ddF technique combines components of Sanger dideoxy sequencing with SSCP. A dideoxy sequencing reaction is performed using one dideoxy terminator and then the reaction products are electrophoresed on nondenaturing polyacrylamide gels to detect alterations in mobility of the termination segments as in SSCP analysis. While ddF is an improvement over SSCP in terms of increased sensitivity, ddF requires the use of expensive dideoxynucleotides and this technique is still limited to the analysis of fragments of the size suitable for SSCP (i.e., fragments of 200-300 bases for optimal detection of mutations).
In addition to the above limitations, all of these methods are limited as to the size of the nucleic acid fragment that can be analyzed. For the direct sequencing approach, sequences of greater than 600 base pairs require cloning, with the consequent delays and expense of either deletion sub-cloning or primer walking, in order b cover the entire fragment.
SSCP and DGGE
have even more severe size limitations. Because of reduced sensitivity to sequence changes, these methods are not considered suitable for larger fragments. Although SSCP
is reportedly able to detect 90 % of single-base substitutions within a 200 base-pair fragment, the detection drops to less than 50 % for 400 base pair fragments. Similarly, the sensitivity of DGGE decreases as the length of the fragment reaches 500 base-pairs. The ddF technique, as a combination of direct sequencing and SSCP, is also limited by the relatively small size of the DNA
that can be screened.
According to a presently preferred embodiment of the present invention the step of searching for any of the nucleic acid sequences described here, in tumor cells or in cells derived from a cancer patient is effected by any suitable technique, including, but not limited to, nucleic acid sequencing, polymerise chain reaction, ligase chain reaction, self sustained synthetic reaction, Q(3-Replicase, cycling probe reaction, branched DNA, restriction fragment length polymorphism analysis, mismatch chemical cleavage, heteroduplex analysis, allele-specific oligonucleotides, denaturing gradient gel electrophoresis, constant denaturant gel electrophoresis, temperature gradient gel electrophoresis and dideoxy fingerprinting.
Detection may also optionally be performed with a chip or other such device.
The nucleic acid sample which includes the candidate region to be analyzed is preferably isolated, amplified and labeled with a reporter group. This reporter group can be a fluorescent group such as phycoerythrin. The labeled nucleic acid is then incubated with the probes immobilized on the chip using a fluidics station. describe the fabrication of fluidics devices and particularly microcapillary devices, in silicon and glass substrates.
Once the reaction is completed, the chip is inserted into a scanner and patterns of hybridization are detected. The hybridization data is collected, as a signal emitted from the reporter groups already incorporated into the nucleic acid, which is now bound to the probes attached to the chip. Since the sequence and position of each probe immobilized on the chip is known, the identity of the nucleic acid hybridized to a given probe can be determined.
It will be appreciated that when utilized along with automated equipment, the above described detection methods can be used to screen multiple samples for a disease and/or pathological condition both rapidly and easily.
Amino acid sequences and peptides The terms "polypeptide," "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an analog or mimetic of a con-esponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers.
Polypeptides can be modified, e.g., by the addition of carbohydrate residues to form glycoproteins. The terms "polypeptide," "peptide" and "protein" include glycoproteins, as well as non-glycoproteins.
Polypeptide products can be biochemically synthesized such as by employing standard solid phase techniques. Such methods include but are not limited to exclusive solid phase synthesis, partial solid phase synthesis methods, fragment condensation, classical solution synthesis. These methods are preferably used when the peptide is relatively short (i.e., 10 kDa) and/or when it cannot be produced by recombinant techniques (i.e., not encoded by a nucleic acid sequence) and therefore involves different chemistry.
Solid phase polypeptide synthesis procedures are well known in the art and further described by John Morrow Stewart and Janis Dillaha Young, Solid Phase Peptide Syntheses (2nd Ed., Pierce Chemical Company, 1984).
Synthetic polypeptides can optionally be purified by preparative high performance liquid chromatography [Creighton T. (1983) Proteins, structures arid molecular principles. WH
Freeman and Co. N.Y.], after which their composition can be confirmed via amino acid sequencing.
In cases where large amounts of a polypeptide are desired, it can be generated using recombinant techniques such as described by Bitter et al., (1987) Methods in Enzymol. 153:516-544, Studier et al. (1990) Methods in Enzymol. 185:60-89, Brisson et al.
(1984) Nature 310:511-514, Takamatsu et al. (1987) EMBO J. 6:307-311, Coruzzi et al. (1984) EMBO J.
3:1671-1680 and Brogli et al., (1984) Science 224:838-843, Gurley et al. (1986) Mol. Cell.
Biol. 6:559-565 and Weissbach & Weissbach, 1988, Methods for Plant Molecular Biology, Academic Press, NY, Section VIII, pp 421-463.
The present invention also encompasses polypeptides encoded by the polynucleotide sequences of the present invention, as well as polypeptides according to the amino acid sequences described herein. The present invention also encompasses homologues of these polypeptides, such homologues can be at least 50 %, at least 55 %, at least 60%, at least 65 %, at least 70 %, at least 75 %, at least 80 %, at least 85 %, at least 95 % or more say 100 homologous to the amino acid sequences set forth below, as can be determined using BlastP
software of the National Center of Biotechnology Information (NCBI) using default parameters, optionally and preferably including the following: filtering on (this option filters repetitive or low-complexity sequences from the query using the Seg (protein) program), scoring matrix is BLOSUM62 for proteins, word size is 3, E value is 10, gap costs are I1, 1 (initialization and extension), and number of alignments shown is 50. Nucleotide (nucleic acid) sequence S homology/identity is preferably determined by using the BlastN software of the National Center of Biotechnology Information (NCBI) using default parameters, which preferably include using the DUST filter program, and also preferably include having an E value of 10, filtering low complexity sequences and a word size of 11. Finally, the present invention also encompasses fragments of the above described polypeptides and polypeptides having mutatio ns, such as deletions, insertions or substitutions of one or more amino acids, either naturally occurring or artificially induced, either randomly or in a targeted fashion.
It will be appreciated that peptides identified according the present invention may be degradation products, synthetic peptides or recombinant peptides as well as peptidomimetics, typically, synthetic peptides and peptoids and semipeptoids which are peptide analogs, which I S may have, for example, modifications rendering the peptides more fable while in a body or more capable of penetrating into cells. Such modifications include, but are not limited to N
terminus modification, C terminus modification, peptide bond modification, including, but not limited to, CH2-NH, CH2-S, CH2-S=O, O=C-NH, CH2-O, CH2-CH2, S=C-NH, CH=CH or CF=CH, backbone modifications, and residue modification. Methods for preparing peptidomimetic compounds are well known in the art and are specified. Further details in this respect are provided hereinunder.
Peptide bonds (-CO-NH-) within the peptide may be substituted, for example, by I~
methylated bonds ( N(CH3)-CO-), ester bonds ( C(R)H-C-O-O-C(R)-N-), ketomethylen bonds (-CO-CH2-), oc-aza bonds (-NH-N(R)-CO-), wherein R is any alkyl, e.g, methyl, carba bonds CH2-NH-), hydroxyethylene bonds (-CH(OH)-CH2-), thioamide bonds (-CS-NH-), olefinic double bonds (-CH=CH-), retro amide bonds (-NH-CO-), peptide derivatives (-N(R)-CH2-CO-), wherein R is the "normal" side chain, naturally presented on the carbon atom.
These modifications can occur at any of the bonds along the peptide chain and even at several (2-3) at the same time.
Natural aromatic amino acids, Trp, Tyr and Phe, may be substituted for synthetic norr natural acid such as Phenylglycine, TIC, naphthylelanine (Nol), ring-methylated derivatives of Phe, halogenated derivatives of Phe or o-methyl-Tyr.
In addition to the above, the peptides of the present invention may also include one or more modified amino acids or one or more norramino acid monomers (e.g. fatty acids, complex carbohydrates etc).
As used herein in the specification and in the claims section below the term "amino acid"
or "amino acids" is understood to include the 20 naturally occurring amino acids; those amino acids often modified post-translationally in vivo, including, for example, hydroxyproline, phosphoserine and phosphothreonine; and other unusual amino acids including, but not limited to, 2-aminoadipic acid, hydroxylysine, isodesmosine, nor-valine, nor-leucine and ornithine.
Furthermore, the term "amino acid" includes both D- and Lramino acids.
Table I non-conventional or modified amino acids which can be used with the present invention.
Table 1 Non-conventional Code Non-conventional Code amino amino acid acid a-aminobutyric Abu LrN-methylalanine Nmala acid a-amino-a-methylbutyrateMgabu >~N-methylarginine Nmarg aminocyclopropane-Cpro IrN-methylasparagineNmasn Carboxylate LrN-methylaspartic Nmasp acid aminoisobutyric Aib LrN-methylcysteine Nmcys acid aminonorbornyl- Norb LrN-methylglutamine Nmgin Carboxylate LrN-methylglutamic Nmglu acid Cyclohexylalanine Chexa IrN-methylhistidine Nmhis CyclopentylalanineCpen LrN-methylisolleucineNmile D-alanine Dal LrN-methylleucine Nmleu D-arginine Darg LrN-methyllysine Nmlys D-aspartic acid Dasp L-N-methylmethionineNmmet D-cysteine Dcys L-N-methylnorleucineNmnle D-glutamine Dgln L-N-methylnorvalineNmova D-glutamic acid Dglu L-N-methylornithineNmorn D-histidine Dhis L-N-methylphenylalanineNmphe D-isoleucine Dile L-N-methylproline Nmpro D-leucine Dleu L-N-methylserine Nmser D-lysine Dlys L-N-methylthreonineNmthr D-methionine Dmet L-N-methyltryptophanNmtrp D-ornithine Dorn L-N-methyltyrosine Nmtyr D-phenylalanine Dphe L-N-methylvaline Nmval D-proline Dpro L-N-methylethylglycineNmetg D-serine Dser IrN-methyl-t-butylglycineNmtbug D-threonine Dthr Lrnorleucine N]e D-tryptophan Dtrp Lrnorvaline Nva D-tyrosine Dtyr a-methyl-aminoisobutyrateMaib D-valine Dval a-methyl-y-aminobutyrateMgabu D-a-methylalanine Dmala a-methylcyclohexylalanineMchexa D-a-methylarginineDmarg a-methylcyclopentylalanineMcpen D-a-methylasparagineDmasn a-methyl-a-napthylalanineManap D-a-methylaspartateDmasp a- methylpenicillamineMpen D-a-methylcysteineDmcys N-(4-aminobutyl)glycineNglu D-a-methylglutamineDmgln N-(2-aminoethyl)glycineNaeg D-a-methylhistidineDmhis N-(3-aminopropyl)glycineNorn D-a-methylisoleucineDmile N- amino-a-methylbutyrateNmaabu D-a-methylleucine Dmleu a-napthylalanine Anap D-a-methyllysine Dmlys N-benzylglycine Nphe D-a-methylmethionineDmmet N-(2-carbamylethyl)glycineNgln D-a-methylornithineDmorn N-(carbamylmethyl)glycineNasn D-a-methylphenylalanineDmphe N-(2-carboxyethyl)glycineNglu D-a-methylprolineDmpro N-(carboxymethyl)glycineNasp D-a-methylserine Dmser N-cyclobutylglycineNcbut D-a-methylthreonineDmthr N-cycloheptylglycineNchep D-a-methyltryptophanDmtrp N-cyclohexylglycineNchex D-a-methyltyrosineDmty N-cyclodecylglycineNedec D-a-methylvaline Dmval N-cyclododeclglycineNcdod D-a-methylalnine Dnmala N-cyclooctylglycineNcoct D-a-methylarginineDnmarg N-cyclopropylglycineNcpro D-a-methylasparagineDnmasn N-cycloundecylglycineNcund D-a-methylasparatateDnmasp N-(2,2-diphenylethyl)glycineNbhm D-a-methylcysteineDnmcys N-(3,3- Nbhe diphenylpropyl)glycine D-N-methylleucineDnmleu N-(3-indolylyethyl)Nhtrp glycine D-N-methyllysine Dnmlys N-methyl-y-aminobutyrateNmgabu N- Nmchexa D-N-methylmethionineDnmmet methylcyclohexylalanine D-N-methylornithineDnmorn N-methylcyclopentylalanineNmcpen N-methylglycine Nala D-N-methylphenylalanineDnmphe N-methylaminoisobutyrateNmaib D-N-methylproline Dnmpro N-(1-methylpropyl)glycineNile D-N-methylserine Dnmser N-(2-methylpropyl)glycineNile D-N-methylserine Dnmser N-(2-methylpropyl)glycineNleu D-N-methylthreonineDnmthr D-N-methyltryptophanDnmtrp N-(1-methylethyl)glycineNva D-N-methyltyrosineDnmtyr N-methyla-napthylalanineNmanap D-N-methylvaline Dnmval N-methylpenicillamineNmpen y-aminobutyric Gabu N-(p-hydroxyphenyl)glycineNhtyr acid Lrt-butylglycine Tbug N-(thiomethyl)glycineNcys Irethylglycine Etg penicillamine Pen L-homophenylalanineHphe L-a-methylalanine Mala L-a-methylarginineMarg L-a-methylasparagineMasn L-a-methylaspartateMasp L-a-methyl-t-butylglycineMtbug L-a-methylcysteineMcys L-methylethylglycineMetg L-a-methylglutamineMgln L-a-methylglutamateMglu L-a-methylhistidineMhis L-a-methylhomo Mhphe phenylalanine L-a-methylisoleucineMile N-(2-methylthioethyl)glycineNmet D-N-methylglutamineDnmgln N-(3- Narg guanidinopropyl)glycine D-N-methylglutamateDnmglu N-(1-hydroxyethyl)glycineNthr D-N-methylhistidineDnmhis N-(hydroxyethyl)glycineNser D-N-methylisoleucineDnmile N-(imidazolylethyl)glycineNhis D-N-methylleucine Dnmleu N-(3-indolylyethyl)glycineNhtrp D-N-methyllysine Dnmlys N-methyl-'y-aminobutyrateNmgabu N- Nmchexa D-N-methylmethionineDnmmet methylcyclohexylalanine D-N-methylornithineDnmorn N-methylcyclopentylalanineNmcpen N-methylglycine Nala D-N-methylphenylalanineDnmphe N-methylaminoisobutyrateNmaib D-N-methylproline Dnmpro N-(1-methylpropyl)glycineNile D-N-methylserine Dnmser N-(2-methylpropyl)glycineNleu D-N-methylthreonineDnmthr D-N-methyltryptophanDnmtrp N-(1-methylethyl)glycineNval D-N-methyltyrosineDnmtyr N-methyla-napthylalanineNmanap D-N-methylvaline Dnmval N-methylpenicillamineNmpen 'y-aminobutyric Gabu N-(p-hydroxyphenyl)glycineNhtyr acid Lrt-butylglycine Tbug N-(thiomethyl)glycineNcys L-ethylglycine Etg penicillamine Pen IrhomophenylalanineHphe Ira-methylalanine Mala L-a-methylarginineMarg L-a-methylasparagineMasn L-a-methylaspartateMasp L-a-methyl-t-butylglycineMtbug L-a-methylcysteineMcys L-methylethylglycineMetg L-a-methylglutamineMgln L-a-methylglutamateMglu L-a-methylhistidineMhis L-a- Mhphe methylhomophenylalanine L-a-methylisoleucineMile N-(2-methylthioethyl)glycineNmet L-a-methylleucineMleu Lra-methyllysine Mlys L-a-methylmethionineMmet L-a-methylnorleucineMnle L-a-methylnorvalineMnva L-a-methylornithineMorn L-a-methylphenylalanineMphe L-a-methylproline Mpro L-a-methylserine mser L-a-methylthreonineMthr Ira-methylvaline Mtrp L-a-methyltyrosine Mtyr Lra-methylleucineMval L-N- Nmhphe Nnbhm methylhomophenylalanine N-(N-(2,2-diphenylethyl) N-(N-(3,3-diphenylpropyl) carbamylmethyl-glycineNnbhm carbamylmethyl(1)glyeineNnbhe 1-carboxy-1-(2,2-diphenylNmbc ethylamino)cyclopropane Table 1 Cont.
Since the peptides of the present invention are preferably utilized in diagnostics which require the peptides to be in soluble form, the peptides of the present invention preferably include one or more norrnatural or natural polar amino acids, including but not limited to serine and threonine which are capable of increasing peptide solubility due to their hydroxyl-containing side chain.
The peptides of the present invention are preferably utilized in a linear form, although it will be appreciated that in cases where cyclicization does not severely interfere with peptide characteristics, cyclic forms of the peptide can also be utilized.
The peptides of present invention can be biochemically synthesized such as by using standard solid phase techniques. These methods include exclusive solid phase synthesis well known in the art, partial solid phase synthesis methods, fragment condensation, classical solution synthesis. These methods are preferably used when the peptide is relatively short (i.e., 10 kDa) and/or when it cannot be produced by recombinant techniques (i.e., not encoded by a nucleic acid sequence) and therefore involves different chemistry.
Synthetic peptides can be purified by preparative high performance liquid chromatography and the composition of which can be confirmed via amino acid sequencing.
In cases where large amounts of the peptides of the present invention are desired, the peptides of the present invention can be generated using recombinant techniques such as described by Bitter et al., (1987) Methods in Enzymol. 153:516-544, Studier et al. (1990) Methods in Enzymol. 185:60-89, Brisson et al. (1984) Nature 310:511-514, Takamatsu et al.
(1987) EMBO J. 6:307-311, Coruzzi et al. (1984) EMBO J. 3:1671-1680 and Brogli et al., (1984) Science 224:838-843, Gurley et al. (1986) Mol. Cell. Biol. 6:559-565 and Weissbach &
Weissbach, 1988, Methods for Plant Molecular Biology, Academic Press, NY, Section VIII, pp 421-463 and also as described above.
Antibodies "Antibody" refers to a polypeptide ligand that is preferably substantially encoded by an immunoglobulin gene or immunoglobulin genes, or fragments thereof, which specifically binds and recognizes an epitope (e.g., an antigen). The recognized immunoglobulin genes include the kappa and lambda light chain constant region genes, the alpha, gamma, delta, epsilon and mu heavy chain constant region genes, and the myriad-immunoglobulin variable region genes.
Antibodies exist, e.g., as intact immunoglobulins or as a number of well characterized fragments produced by digestion with various peptidases. This includes, e.g., Fab' and F(ab)'z fragments.
The term "antibody," as used herein, also includes antibody fragments either produced by the modification of whole antibodies or those synthesized de novo using recombinant DNA
methodologies. It also includes polyclonal antibodies, monoclonal antibodies, chimeric antibodies, humanized antibodies, or single chain antibodies. "Fc" portion of an antibody refers to that portion of an immunoglobulin heavy chain that comprises one or more heavy chain constant region domains, CHI, CH2 and CH3, but does not include the heavy chain variable region.
The functional fragments of antibodies, such as Fab, F(ab')2, and Fv that are capable of binding to macrophages, are described as follows: (1) Fab, the fragment which contains a monovalent antigen-binding fragment of an antibody molecule, can be produced by digestion of whole antibody with the enzyme papain to yield an intact light chain and a portion of one heavy chain; (2) Fab', the, fragment of an antibody molecule that can be obtained by treating whole antibody with pepsin, followed by reduction, to yield an intact light chain and a portion of the heavy chain; two Fab' fragments are obtained per antibody molecule; (3) (Fab')2, the fragment of the antibody that can be obtained by treating whole antibody with the enzyme pepsin without subsequent reduction; F(ab')2 is a dimer of two Fab' fragments held together by two disulfide bonds; (4) Fv, defined as a genetically engineered fragment containing the variable region of the light chain and the variable region of the heavy chain expressed as two chains; and (5) Single chain antibody ("SCA"), a genetically engineered molecule containing the variable region of the light chain and the variable region of the heavy chain, linked by a suitable polypeptide linker as a genetically fused single chain molecule.
Methods of producing polyclonal and monoclonal antibodies as well as fragments thereof are well known in the art (See for example, Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York, 1988, incorporated herein by reference).
Antibody fragments according to the present invention can be prepared by proteolytic hydrolysis of the antibody or by expression in E. coli or mammalian cells (e.g. Chinese hamster ovary cell culture or other protein expression systems) of DNA encoding the fragment.
Antibody fragments can be obtained by pepsin or papain digestion of whole antibodies by conventional methods. For example, antibody fragments can be produced by enzymatic cleavage of antibodies with pepsin to provide a SS fragment denoted F(ab')2.
This fragment can be further cleaved using a thiol reducing agent, and optionally a blocking group for the sulfhydryl groups resulting from cleavage of disulfide linkages, to produce 3.SS Fab' monovalent fragments. Alternatively, an enzymatic cleavage using pepsin produces two monovalent Fab' fragments and an Fc fragment directly. These methods are described, for example, by Goldenberg, U.S. Pat. Nos. 4,036,945 and 4,331,647, and references contained therein, which patents are hereby incorporated by reference in their entirety.
See also Porter, R.
R. [Biochem. J. 73: 1 19-126 ( 1959)]. Other methods of cleaving antibodies, such as separation of heavy chains to form monovalent light-heavy chain fragments, further cleavage of fragments, or other enzymatic, chemical, or genetic techniques may also be used, so long as the fragments bind to the antigen that is recognized by the intact antibody.
Fv fragments comprise an association of VH and VL chains. This association may be noncovalent, as described in hbar et al. [Proc. Nat'1 Acad. Sci. USA 69:2659-62 (19720].
Alternatively, the variable chains can be linked by an intermolecular disulfide bond or cross-linked by chemicals such as glutaraldehyde. Preferably, the Fv fragments comprise VH and VL
chains connected by a peptide linker. These single-chain antigen binding proteins (sFv) are prepared by constructing a structural gene comprising DNA sequences encoding the VH and VL
domains connected by an oligonucleotide. The structural gene is inserted into an expression vector, which is subsequently introduced into a host cell such as E. coli. The recombinant host cells synthesize a single polypeptide chain with a linker peptide bridging the two V domains.
Methods for producing sFvs are described, for example, by [Whitlow and Filpula, Methods 2:
97-105 (1991); Bird et al., Science 242:423-426 (1988); Pack et al., Bio/Technology 11:1271-77 (1993); and U.S. Pat. No. 4,946,778, which is hereby incorporated by reference in its entirety.
Another form of an antibody fragment is a peptide coding for a single complementarit~
determining region (CDR). CDR peptides ("minimal recognition units") can be obtained by constructing genes encoding the CDR of an antibody of interest. Such genes are prepared, for example, by using the polymerise chain reaction to synthesize the variable region from RNA of antibody producing cells. See, for example, Larrick and Fry [Methods, 2: 106-10 (1991)].
Humanized forms of non-human (e.g., murine) antibodies are chimeric molecules of immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab') or other antigen-binding subsequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin. Humanized antibodies include human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a norrhuman species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity. In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding norrhuman residues. Humanized antibodies may also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin [Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol., 2:593-596 (1992)].
Methods for humanizing norrhuman antibodies are well known in the art.
Generally, a humanized antibody has one or more amino acid residues introduced into it from a source which is norrhuman. These non-human amino acid residues are often referred to as import residues, which are typically taken from an import variable domain. Humanization can be essentially performed following the method of Winter and co-workers [Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)], by substituting rodent CDRs or CDR sequences for the corresponding sequences IS of a human antibody. Accordingly, such humanized antibodies are chimeric antibodies (U.S.
Pat. No. 4,816,567), wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR
residues are substituted by residues from analogous sites in rodent antibodies.
Human antibodies can also be produced using various techniques known in the art, including phage display libraries [Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991);
Marks et al., J. Mol. Biol., 222:581 (1991)]. The techniques of Cole et al.
and Boerner et al. are also available for the preparation of human monoclonal antibodies (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985) and Boerner et al., J. Immunol., 147(1):86-95 (1991)]. Similarly, human antibodies can be made by introduction of human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126;
5,633,425;
5,661,016, and in the following scientific publications: Marks et al., Bio/Technology 10,: 779-783 ( I 992); Lonberg et al., Nature 368: 856-859 ( 1994); Morrison, Nature 368 812- I 3 ( 1994);
Fishwild et al., Nature Biotechnology 14, 845-51 (1996); Neuberger, Nature Biotechnology 14:
826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13, 65-93 (1995).
Preferably, the antibody of this aspect of the present invention specifically binds at least one epitope of the polypeptide variants of the present invention. As used herein, the term "epitope" refers to any antigenic determinant on an antigen to which the paratope of an antibody binds.
Epitopic determinants usually consist of chemically active surface groupings of molecules such as amino acids or carbohydrate side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics.
Optionally, a unique epitope may be created in a variant due to a change in one or more post-translational modifications, including but not limited to glycosylation and/or phosphorylation, as described below. Such a change may also cause a new epitope to be created, for example through removal of glycosylation at a particular site.
An epitope according to the present invention may also optionally comprise part or all of a unique sequence portion of a variant according to the present invention in combination with at least one other portion of the variant which is not contiguous to the unique sequence portion in the linear polypeptide itself, yet which are able to form an epitope in combination. One or more unique sequence portions may optionally combine with one or more other noncontiguous portions of the variant (including a portion which may have high homology to a portion of the known protein) to form an epitope.
Immunoassays In another embodiment of the present invention, an immunoassay can be used to qualitatively or quantitatively detect and analyze markers in a sample. This method comprises:
providing an antibody that specifically binds to a marker; contacting a sample with the antibody;
and detecting the presence of a complex of the antibody bound to the marker in the sample.
To prepare an antibody that specifically binds to a marker, purified protein markers can be used. Antibodies that specifically bind to a protein marker can be prepared using any suitable methods known in the art.
After the antibody is provided, a marker can be detected and/or quantified using any of a number of well recognized immunological binding assays. Useful assays include, for example, an enzyme immune assay (EIA) such as enzyme-linked immunosorbent assay (ELISA), a radioimmune assay (RIA), a Western blot assay, or a slot blot assay see, e.g., U.S. Pat. Nos.
4,366,241; 4,376,110; 4,517,288; and 4,837,168). Generally, a sample obtained from a subject can be contacted with the antibody that specifically binds the marker.
Optionally, the antibody can be fixed to a solid support to facilitate washing and subsequent isolation of the complex, prior to contacting the antibody with a sample. Examples of solid supports include but are not limited to glass or plastic in the form of, e.g., a microtiter plate, a stick, a bead, or a microbead. Antibodies can also be attached to a solid support.
After incubating the sample with antibodies, the mixture is washed and the antibody marker complex formed can be detected. This can be accomplished by incubating the washed mixture with a detection reagent. Alternatively, the marker in the sample can be detected using an indirect assay, wherein, for example, a second, labeled antibody is used to detect bound marker-specific antibody, and/or in a competition or inhibition assay wherein, for example, a monoclonal antibody which binds to a distinct epitope of the marker are incubated simultaneously with the mixture.
Throughout the assays, incubation and/or washing steps may be required after each combination of reagents. Incubation steps can vary from about 5 seconds to several hours, preferably from about 5 minutes to about 24 hours. However, the incubation time will depend upon the assay format, marker, volume of solution, concentrations and the like. Usually the assays will be earned out at ambient temperature, although they can be conducted over a range of temperatures, such as 10 °C to 40 °C.
The immunoassay can be used to determine a test amount of a marker in a sample from a subject. First, a test amount of a marker in a sample can be detected using the immunoassay methods described above. If a marker is present in the sample, it will form an antibody-marker complex with an antibody that specifically binds the marker under suitable incubation conditions described above. The amount of an antibody marker complex can optionally be determined by comparing to a standard. As noted above, the test amount of marker need not be measured in absolute units, as long as the unit of measurement can be compared to a control amount and/or signal.
Preferably used are antibodies which specifically interact with the polypeptides of the present invention and not with wild type proteins or other isoforms thereof, for example. Such antibodies are directed, for example, to the unique sequence portions of the polypeptide variants of the present invention, including but not limited to bridges, heads, tails and insertions described in greater detail below. Preferred embodiments of antibodies according to the present invention are described in greater detail with regard to the section entitled "Antibodies".
Radio-immunoassay (RIA): In one version, this method involves precipitation of the desired substrate and in the methods detailed hereinbelow, with a specific antibody and radiolabelled antibody binding protein (e.g., protein A labeled with 1125) immobilized on a precipitable earner such as agarose beads. The number of counts in the precipitated pellet is proportional to the amount of substrate.
In an alternate version of the RIA, a labeled substrate and an unlabelled antibody binding protein are employed. A sample containing an unknown amount of substrate is added in varying amounts. The decrease in precipitated counts from the labeled substrate 's proportional to the amount of substrate in the added sample.
Enzyme linked immunosorbent assay (ELISA): This method involves fixation of a sample (e.g., fixed cells or a proteinaceous solution) containing a protein substrate to a surface such as a well of a microtiter plate. A substrate specific antibody coupled to an enzyme is applied and allowed to bind to the substrate. Presence of the antibody is then detected and quantitated by a colorimetric reaction employing the enzyme coupled to the antibody. Enzymes commonly employed in this method include horseradish peroxidase and alkaline phosphatase. If well calibrated and within the linear range of response, the amount of substrate present in the sample is proportional to the amount of color produced. A substrate standard is generally employed to improve quantitative accuracy.
Western blot: This method involves separation of a substrate from other protein by means of an acrylamide gel followed by transfer of the substrate to a membrane (e.g., nylon or PVDF).
Presence of the substrate is then detected by antibodies specific to the substrate, which are in turn detected by antibody binding reagents. Antibody binding reagents may be, for example, protein A, or other antibodies. Antibody binding reagents may be radiolabelled or enzyme linked as described hereinabove. Detection may be by autoradiography, colorimetric reaction or chemiluminescence. This method allows both quantitation of an amount of substrate and determination of its identity by a relative position on the membrane which is indicative of a migration distance in the acrylamide gel during electrophoresis.
Irnmunohistochemical analysis: This method involves detection of a substrate in situ in fixed cells by substrate specific antibodies. The substrate specific antibodies may be enzyme linked or linked to fluorophores. Detection is by microscopy and subjective evaluation. If enzyme linked antibodies are employed, a colorimetric reaction may be required.
Fluorescence activated cell sorting (FRCS): This method involves detection of a substrate in situ in cells by substrate specific antibodies. The substrate specific antibodies are linked to fluorophores. Detection is by means of a cell sorting machine which reads the wavelength of light emitted from each cell as it passes through a light beam.
This method may employ two or more antibodies simultaneously.
1 S Radio-imaging Methods These methods include but are not limited to, positron emission tomography (PET) single photon emission computed bmography (SPELT). Both of these techniques are norr invasive, and can be used to detect and/or measure a wide variety of tissue events and/or functions, such as detecting cancerous cells for example. Unlike PET, SPELT
can optionally be used with two labels simultaneously. SPELT has some other advantages as well, for example with regard to cost and the types of labels that can be used. For example, US
Patent No.
6,696,686 describes the use of SPELT for detection of breast cancer, and is hereby incorporated by reference as if fully set forth herein.
Display Libraries According to still another aspect of the present invention there is provided a display library comprising a plurality of display vehicles (such as phages, viruses or bacteria) each displaying at least 6, at least 7, at least 8, at least 9, at least 10, 10-15, 12-17, 15-20, 15-30 or 20-50 consecutive amino acids derived from the polypeptide sequences of the present invention.
Methods of constructing such display libraries are well known in the art. Such methods are described in, for example, Young AC, et al., "The three-dimensional structures of a polysaccharide binding antibody to Cryptococcus neoformans and its complex with a peptide from a phage display library: implications for the identification of peptide mimotopes" J Mol Biol 1997 Dec 12;274(4):622-34; Giebel LB et al. "Screening of cyclic peptide phage libraries identifies ligands that bind streptavidin with high affinities" Biochemistry 1995 Nov 28;34(47):15430-5; Davies EL et al., "Selection of specific phage-display antibodies using libraries derived from chicken immunoglobulin genes" J Immunol Methods 1995 Oct 12;186( 1 ):125-35; Jones C RT al. "Current trends in molecular recognition and bioseparation" J
Chromatogr A 1995 Jul 14;707( 1 ):3-22; Deng SJ et al. "Basis for selection of improved carbohydrate-binding single-chain antibodies from synthetic gene libraries"
Proc Natl Acad Sci U S A 1995 May 23;92(11):4992-6; and Deng SJ et al. "Selection of antibody single-chain variable fragments with improved carbohydrate binding by phage display" J Biol Chem 1994 Apr 1;269(13):9533-8, which are incorporated herein by reference.
The following sections relate to Candidate Marker Examples (first section) and to Experimental Data for these Marker Examples (second section). It should be noted that Table numbering is restarted within each section.
CANDIDATE MARKER EXAMPLES SECTION
This Section relates to Examples of sequences according to the present invention, including illustrative methods of selection thereof.
Description of the methodology undertaken to uncover the biomolecular sequences of the present invention Human ESTs and cDNAs were obtained from GenBank versions 136 (June 15, 2003 ftp.ncbi.nih.gov/genbank/release.notes/gb136.release.notes); NCBI genome assembly of April 2003; Refseq sequences from June 2003; Genbank version 139 (December 2003);
Human Genome from NCBI (Build 34) (from Oct 2003); and Ref~eq sequences from December 2003;
and from the LifeSeq library of Incyte Corporation (ES Ts only; Wilmington, DE, USA). With regard to GenBank sequences, the human EST sequences from the EST (GBEST) section and the human mRNA sequences from the primate (GBPRI) section were used; also the human nucleotide Refseq mRNA sequences were used (see for example www.ncbi.nlm.nih.gov/Genbank/GenbankOverview.html and for a reference to the EST section, see www.ncbi.nlm.nih.gov/dbEST/; a general reference to dbEST, the EST
database in GenBank, may be found in Boguski et al, Nat Genet. 1993 Aug;4(4):332-3; all of which are hereby incorporated by reference as if fully set forth herein).
Novel splice variants were predicted using the LEADS clustering and assembly system as described in Sorek, R., Ast, G. & Graur, D. Alu-containing exons are alternatively spliced.
Genome Res 12, 1060-7 (2002); US patent No: 6,625,545; and U.S. Pat. Appl. No.
10/426,002, published as US20040101876 on May 27 2004; all of which are hereby incorporated by reference as if fully set forth herein. Briefly, the software cleans the expressed sequences from repeats, vectors and immunoglobulins. It then aligns the expressed sequences to the genome taking alternatively splicing into account and clusters overlapping expressed sequences into "clusters" that represent genes or partial genes.
These were annotated using the GeneCarta (Compugen, Tel-Aviv, Israel) platform. The GeneCarta platform includes a rich pool of annotations, sequence information (particularly of spliced sequences), chromosomal information, alignments, and additional information such as SNPs, gene ontology terms, expression profiles, functional analyses, detailed domain structures, known and predicted proteins and detailed homology reports.
A brief explanation is provided with regard to the method of selecting the candidates.
However, it should noted that this explanation is provided for descriptive purposes only, and is not intended to be limiting in any way. The potential markers were identified by a computational process that was designed to find genes and/or their splice variants that are over-expressed in tumor tissues, by using databases of expressed sequences. Various parameters related to the information in the EST libraries, determined according to a manual classification process, were used to assist in locating genes and/or splice variants thereof that are over-expressed in cancerous tissues. The detailed description of the selection method is presented in Example 1 below. The cancer biomarkers selection engine and the following wet validation stages are schematically summarized in Figure 1.
Identification of differentially expressed gene products - Algorithm In order to distinguish between differentially expressed gene products and constitutively expressed genes (i.e., house keeping genes ) an algorithm based on an analysis of frequencies was configured. A specific algorithm for identification of transcripts over expressed in cancer is described hereinbelow.
Dry analysis Library annotation - EST libraries are manually classified according to:
(i) Tissue origin (ii) Biological source - Examples of frequently used biological sources for construction of EST libraries include cancer cell-lines; normal tissues; cancer tissues; fetal tissues; and others such as normal cell lines and pools of normal cell-lines, cancer cell-lines and combinations thereof. . A specific description of abbreviations used below with regard to these tissues/cell lines etc is given above.
(iii) Protocol of library construction - various methods are known in the art for library construction including normalized library construction;
non-normalized library construction; subtracted libraries; ORESTES and others. It will be appreciated that at times the protocol of library construction is not indicated.
The following rules were followed:
EST libraries originating from identical biological samples are considered as a single library.
EST libraries which included above-average levels of contamination, such as DNA
contamination for example, were eliminated. The presence of such contamination was determined as follows. For each library, the number of unspliced ESTs that are not fully contained within other spliced sequences was counted. If the percentage of such sequences (as compared to all other sequences) was at least 4 standard deviations above the average for all libraries being analyzed, this library was tagged as being contaminated and was eliminated from further consideration in the below analysis (see also Sorek, R. & Safer, H.M. A novel algorithm for computational identification of contaminated EST libraries. Nucleic Acids Res 31, 1067-74 (2003)for further details).
Clusters (genes) having at least five sequences including at least two sequences from the tissue of interest were analyzed. Splice variants were identified by using the LEADS software package as described above.
Identification of genes over expressed in cancer.
Two different scoring algorithms were developed.
Libraries score -candidate sequences which are supported by a number of cancer libraries, are more likely to serve as specific and effective diagnostic markers.
The basic algorithm - for each cluster the number of cancer and normal libraries contributing sequences to the cluster was counted. Fisher exact test was used to check if cancer libraries are significantly over-represented in the cluster as compared to the total number of cancer and normal libraries.
Library counting: Small libraries (e.g., less than 1000 sequences) were excluded from consideration unless they participate in the cluster. For this reason, the total number of libraries is actually adjusted for each cluster.
Clones no. score - Generally, when the number of ESTs is much higher in the cancer libraries relative to the normal libraries it might indicate actual over-expression.
The algorithm -Clone counting : For counting EST clones each library protocol class was given a weight based on our belief of how much the protocol reflects actual expression levels:
(i) non-normalized : 1 (ii) normalized : 0.2 (iii) all other classes : 0.1 Clones number score - The total weighted number of EST clones from cancer libraries was compared to the EST clones from normal libraries. To avoid cases where one library contributes to the majority of the score, the contribution of the library that gives most clones for a given cluster was limited to 2 clones.
The score was computed as C+1 C
n+1 N
where:
c - weighted number of "cancer" clones in the cluster.
C- weighted number of clones in all "cancer" libraries.
n - weighted number of "normal" clones in the cluster.
N- weighted number of clones in all "normal" libraries.
Clones number score significance - Fisher exact test was used to check if EST
clones from cancer libraries are significantly over-represented in the cluster as compared to the total number of EST clones from cancer and normal libraries.
Two search approaches were used to find either general cancer-specific candidates or tumor specific candidates.
~ Libraries/sequences originating from tumor tissues are counted as well as libraries originating from cancer cell-lines ("normal" cell-lines were ignored).
~ Only libraries/sequences originating from tumor tissues are counted Identification of tissue specific genes For detection of tissue specific clusters, tissue libraries/sequences mere compared to the total number of libraries/sequences in cluster. Similar statistical tools to those described in above were employed to identify tissue specific genes. Tissue abbreviations are the same as for cancerous tissues, but are indicated with the header "normal tissue".
The algorithm - for each tested tissue T and for each tested cluster the following were examined:
1. Each cluster includes at least 2 libraries from the tissue T. At least 3 clones (weighed - as described above) from tissue T in the cluster; and 2. Clones from the tissue T are at least 40 % from all the clones participating in the tested cluster Fisher exact test Rvalues were computed both for library and weighted clone counts to check that the counts are statistically significant.
Identification of splice variants over expressed in cancer of clusters which are not over expressed in cancer Cancer-specific splice variants containing a unique region were identified.
Identification of unique sequence regions in splice variants A Region is defined as a group of adjacent exons that always appear or do not appear together in each splice variant.
A "segment" (sometimes referred also as "seg" or "node") is defined as the shortest contiguous transcribed region without known splicing inside.
, Only reliable ESTs were considered for region and segment analysis. An EST
was defined as unreliable i~
(i) Unspliced;
(ii) Not covered by RNA;
(iii) Not covered by spliced ESTs; and (iv) Alignment to the genome ends in proximity of long poly-A stretch or starts in proximity of long poly-T stretch.
Only reliable regions were selected for further scoring. Unique sequence regions were considered reliable i~
(i) Aligned to the genome; and (ii) Regions supported by more than 2 ESTs.
The algorithm Each unique sequence region divides the set of transcripts into 2 groups:
(i) Transcripts containing this region (group TA).
(ii) Transcripts not containing this region (group TB).
The set of EST clones of every cluster is divided into 3 groups:
(i) Supporting (originating from) transcripts of group TA (S 1 ).
(ii) Supporting transcripts of group TB (S2).
(iii) Supporting transcripts from both groups (S3).
Library and clones number scores described above were given to S1 group.
Fisher Exact Test P-values were used to check i~
S 1 is significantly enriched by cancer EST clones compared to S2; and S1 is significantly enriched by cancer EST clones compared to cluster background (S1+S2+S3).
Identification of unique sequence regions and division of the group of transcripts accordingly is illustrated in Figure 2. Each of these unique sequence regions corresponds to a segment, also termed herein a "node".
Region 1: conunon to all transcripts, thus it is preferably not considered for determining differential expression between variants; Region 2: specific to Transcript 1;
Region 3: specific to Transcripts 2+3; Region 4: specific to Transcript 3; Region 5: specific to Transcripts 1 and 2;
Region 6: specific to Transcript 1.
Identification of cancer specific splice variants of genes over expressed in cancer A search for EST supported (no mRNA) regions for genes o~
(i) known cancer markers (ii) Genes shown to be over-expressed in cancer in published micro-array experiments.
Reliable EST supported-regions were defined as supported by minimum of one of the following:
(i) 3 spliced ESTs; or (ii) 2 spliced ESTs from 2 libraries;
(iii) 10 unspliced ESTs from 2 libraries, or (iv) 3libraries.
Actual Marker Examples The following examples relate to specific actual marker examples. It should be noted that Table numbering is restarted within each example related to a particular Cluster, as indicated by the titles below.
EXPERIMENTAL EXAMPLES SECTION
This Section relates to Examples describing experiments involving these sequences, and illustrative, norrlimiting examples of methods, assays and uses thereof. The materials and experimental procedures are explained first, as all experiments used them as a basis for the work I O that was performed.
The markers of the present invention were tested with regard to their expression in various cancerous and norrcancerous tissue samples. A description of the samples used in the panel is provided in Table 1 below. A description of the samples used in the normal tissue panel is provided in Table 2 below. Tests were then performed as described in the "Materials and Experimental Procedures" section below.
Table 1: Tissue samples in testing panel Lot Sample numbe Gra name r ource Tissue Pathology de gender/age 2-A-Pap ILS- Papillary Adeno G2 1408 BS ovary adenocarcinoma 2 53/F
3-A-Pap ILS- Papillary Adeno G2 1431 BS ovary adenocarcinoma 2 52/F
4-A-Pap CystAdeno ILS- Papillary G2 7286 BS ovary cystadenocarcinoma2 50/F
1-A-Pap ILS- BS ovary Papillary 3 73/F
Adeno G3 1406 adenocarcinoma 14-B-AdenoA50111BioChai G2 1 n ovary Adenocarcinoma 2 41/F
5-G-Adeno 99-12- Adenocarcinoma G3 6432 GOG ovary (Stage3C) 3 46/F
6-A-Adeno A0106 BS ovary adenocarcinoma 3 51/F
IND-7-A-Adeno 00375 BS ovary adenocarcinoma 3 59/F
A50111BioChai 8-B-Adeno 3 n ovary adenocarcinoma 3 60/F
The detection of hybrid duplexes can be carried out by a number of methods.
Typically, hybridization duplexes are separated from unhybridized nucleic acids and the labels bound to the duplexes are then detected. Such labels refer to radioactive, fluorescent, biological or enzymatic tags or labels of standard use in the art. A label can be conjugated to either the oligonucleotide probes or the nucleic acids derived from the biological sample.
Probes can be labeled according to numerous well known methods. Non-limiting examples of radioactive labels include 3H, 14C, 32P, and 35S. Norrlimiting examples of detectable markers include ligands, fluorophores, chemiluminescent agents, enzymes, and antibodies. Other detectable markers for use with probes, which can enable an increase in sensitivity of the method of the invention, include biotin and radio-nucleotides. It will become evident to the person of ordinary skill that the choice of a particular label dictates the manner in which it is bound to the probe.
For example, oligonucleotides of the present invention can be labeled subsequent to synthesis, by incorporating biotinylated dNTPs or rNTP, or some similar means (e.g., photo cross-linking a psoralen derivative of biotin to RNAs), followed by addition of labeled streptavidin (e.g., phycoerythrin-conjugated streptavidin) or the equivalent.
Alternatively, when fluorescently-labeled oligonucleotide probes are used, fluorescein, lissamine, phycoerythrin, rhodamine (Perkin Elmer Cetus), Cy2, Cy3, Cy3.5, CyS, Cy5.5, Cy7, FIuorX
(Amersham) and others [e.g., Kricka et al. (1992), Academic Press San Diego, CalifJ can be attached to the oligonucleotides.
Those skilled in the art will appreciate that wash steps may be employed to wash away excess target DNA or probe as well as unbound conjugate. Further, standard heterogeneous assay formats are suitable for detecting the hybrids using the labels present on the oligonucleotide primers and probes.
It will be appreciated that a variety of controls may be usefully employed to improve accuracy of hybridization assays. For instance, samples may be hybridized to an irrelevant probe and treated with RNAse A prior to hybridization, to assess false hybridization.
Although the present invention is not specifically dependent on the use of a label for the detection of a particular nucleic acid sequence, such a label might be beneficial, by increasing the sensitivity of the detection. Furthermore, it enables automation. Probes can be labeled according to numerous well known methods.
As commonly known, radioactive nucleotides can be incorporated into probes of the invention by several methods. Norr limiting examples of radioactive labels include 3H, '4C, 32P, and 35S.
Those skilled in the art will appreciate that wash steps may be employed to wash away excess target DNA or probe as well as unbound conjugate. Further, standard hetero~neous assay formats are suitable for detecting the hybrids using the labels present on the oligonucleotide primers and probes.
It will be appreciated that a variety of controls may be usefully employed to improve accuracy of hybridization assays.
Probes of the invention can be utilized with naturally occurnng sugar-phosphate backbones as well as modified backbones including phosphorothioates, dithionates, alkyl phosphonates and a-nucleotides and the like. Probes of the invention can be constructed of either ribonucleic acid (RNA) or deoxyribonucleic acid (DNA), and preferably of DNA.
NAT Assays Detection of a nucleic acid of interest in a biological sample may also optionally be effected by NAT-based assays, which involve nucleic acid amplification ~chnology, such as PCR for example (or variations thereof such as real-time PCR for example).
As used herein, a "primer" defines an oligonucleotide which is capable of annealing to (hybridizing with) a target sequence, thereby creating a double stranded region which can serve as an initiation point for DNA synthesis under suitable conditions.
Amplification of a selected, or target, nucleic acid sequence may be carried out by a number of suitable methods. See generally Kwoh et al., 1990, Am. Biotechnol.
Lab. 8:14 Numerous amplification techniques have been described and can be readily adapted to suit particular needs of a person of ordinary skill. Norr limiting examples of amplification techniques include polymerase chain reaction (PCR), ligase chain reaction (LCR), strand displacement amplification (SDA), transcriptiorrbased amplification, the q3 replicase system and NASBA
(Kwoh et al., 1989, Proc. NatI. Acad. Sci. USA 86, 1173-1177; Lizardi et al., 1988, BioTechnology 6:1197-1202; Malek et al., 1994, Methods Mol. Biol., 28:253-260;
and Sambrook et al., 1989, supra).
The terminology "amplification pair" (or "primer pair") refers herein to a pair of oligonucleotides (oligos) of the present invention, which are selected to be used together in amplifying a selected nucleic acid sequence by one of a number of types of amplification processes, preferably a polymerase chain reaction. Other types of amplification processes include ligase chain reaction, strand displacement amplification, or nucleic acid sequence-based amplification, as explained in greater detail below. As commonly known in the art, the oligos are designed to bind to a complementary sequence under selected conditions.
In one particular embodiment, amplification of a nucleic acid sample from a patient is amplified under conditions which favor the amplification of the most abundant differentially expressed nucleic acid. In one preferred embodiment, RT PCR is carried out on an mRNA
sample from a patient under conditions which favor the amplification of the most abundant mRNA. In another preferred embodiment, the amplification of the differentially expressed nucleic acids is carried out simultaneously. It will be realized by a person skilled in the art that such methods could be adapted for the detection of differentially expressed proteins instead of differentially expressed nucleic acid sequences.
The nucleic acid (i.e. DNA or RNA) for practicing the present invention may be obtained according to well known methods.
Oligonucleotide primers of the present invention may be of any suitable length, depending on the particular assay format and the particular needs and targeted genomes employed. Optionally, the oligonucleotide primers are at least 12 nucleotides in length, preferably between 15 and 24 molecules, and they may be adapted to be especially suited to a chosen nucleic acid amplification system. As commonly known in the art, the oligonucleotide primers can be designed by taking into consideration the melting point of hybridization thereof with its targeted sequence (Sambrook et al., 1989, Molecular Cloning -A
Laboratory Manual, 2nd Edition, CSH Laboratories; Ausubel et al., 1989, in Current Protocols in Molecular Biology, John Wiley & Sons Inc., N.Y.).
It will be appreciated that antisense oligonucleotides may be employed to quantify expression of a splice isoform of interest. Such detection is effected at the pre-mRNA level.
Essentially the ability to quantitate transcription from a splice site of interest can be effected based on splice site accessibility. Oligonucleotides may compete with splicing factors for the splice site sequences. Thus, low activity of the antisense oligonucleotide is indicative of splicing activity.
The polymerise chain reaction and other nucleic acid amplification reactions are well known in the art (various norr limiting examples of these reactions are described in greater detail below). The pair of oligonucleotides according to this aspect of the present invention are preferably selected to have compatible melting temperatures (Tm), e.g., melting temperatures which differ by less than that 7 °C, preferably less than 5 °C, more preferably less than 4 °C, most preferably less than 3 °C, ideally between 3 °C and 0 °C.
Polymerise Chain Reaction (PCR): The polymerise chain reaction (PCR), as described in U.S. Pat. Nos. 4,683,195 and 4,683,202 to Mullis and Mullis et al., is a method of increasing the concentration of a segment of target sequence in a mixture of genomic DNA
without cloning or purification. This technology provides one approach to the problems of low target sequence concentration. PCR can be used to directly increase the concentration of the target to an easily detectable level. This process for amplifying the target sequence involves the introduction of a molar excess of two oligonucleotide primers which are complementary to their respective strands of the double-stranded target sequence to the DNA mixture containing the desired target sequence. The mixture is denatured and then allowed to hybridize. Following hybridization, the primers are extended with polymerise so as to form complementary strands. The steps of denaturation, hybridization (annealing), and polymerise extension (elongation) can be repeated as often as needed, in order to obtain relatively high concentrations of a segment of the desired target sequence.
The length of the segment of the desired target sequence is determined by the relative positions of the primers with respect to each other, and, therefore, this length is a controllable parameter. Because the desired segments of the target sequence become the dominant sequences (in terms of concentration) in the mixture, they are said to be "PCR-amplified."
Ligase Chain Reaction (LCR or LAR): The ligase chain reaction [LCR; sometimes referred to as "Ligase Amplification Reaction" (LAR)] has developed into a well-recognized alternative method of amplifying nucleic acids. In LCR, four oligonucleotides, two adjacent oligonucleotides which uniquely hybridize to one strand of target DNA, and a complementary set of adjacent oligonucleotides, which hybridize to the opposite strand are mixed and DNA ligase is added to the mixture. Provided that there is complete complementarity at the junction, ligase will covalently link each set of hybridized molecules. Importantly, in LCR, two probes are ligated together only when they base-pair with sequences in the target sample, without gaps or mismatches. Repeated cycles of denaturation, and ligation amplify a short segment of DNA.
LCR has also been used in combination with PCR to achieve enhanced detection of single-base changes: see for example Segev, PCT Publication No. W09001069 A1 (1990).
However, because the four oligonucleotides used in this assay can pair to form two short ligatable fragments, there is the potential for the generation of target independent background signal. The use of LCR for mutant screening is limited to the examination of specific nucleic acid positions.
Self-Sustained Synthetic Reaction (3SRlNASBA): The self sustained sequence replication reaction (3SR) is a transeriptiorrbased in vitro amplification system that can exponentially amplify RNA sequences at a uniform temperature. The amplified RNA can then be utilized for mutation detection. In this method, an oligonucleotide primer is used to add a phage RNA
polymerise promoter to the 5' end of the sequence of interest. In a cocktail of enzymes and substrates that includes a second primer, reverse transcriptase, RNase H, RNA
polymerise and ribo-and deoxyribonucleoside triphosphates, the target sequence undergoes repeated rounds of transcription, cDNA synthesis and second-strand synthesis to amplify the area of interest. The use of 3SR to detect mutations is kinetically limited to screening small segments of DNA (e.g., 200-300 base pairs).
Q-Beta (Q(3) Replicase: In this method, a probe which recognizes the sequence of interest is attached to the replicatable RNA template for Q[3 replicase. A
previously identified major problem with false positives resulting from the replication of unhybridized probes has been addressed through use of a sequence-specific ligation step. However, available thermostable DNA ligases are not effective on this RNA substrate, so the ligation must be performed by T4 DNA ligase at low temperatures (37 degrees C.). This prevents the use of high temperature as a means of achieving specificity as in the LCR, the ligation event can be used to detect a mutation at the junction site, but not elsewhere.
A successful diagnostic method must be very specific. A straight-forward method of controlling the specificity of nucleic acid hybridization is by controlling the temperature of the reaction. While the 3SR/NASBA, and Q(3 systems are all able to generate a large quantity of signal, one or more of the enzymes involved in each cannot be used at high temperature (i.e., >
55 degrees C). Therefore the reaction temperatures cannot be raised to prevent non-specific hybridization of the probes. If probes are shortened in order to make them melt more easily at low temperatures, the likelihood of having more than one perfect match in a complex genome increases. For these reasons, PCR and LCR currently dominate the research field in detection technologies.
The basis of the amplification procedure in the PCR and LCR is the fact that the products of one cycle become usable templates in all subsequent cycles, consequently doubling the population with each cycle. The final yield of any such doubling system can be expressed as:
(1+X)n ~, where "X" is the mean efficiency (percent copied in each cycle), "n"
is the number of cycles, and "y" is the overall efficiency, or yield of the reaction. If every copy of a target DNA is utilized as a template in every cycle of a polymerase chain reaction, then the mean efficiency is 100 %. If 20 cycles of PCR are performed, then the yield will be 220, or 1,048,576 copies of the starting material. If the reaction conditions reduce the mean efficiency to 85 %, then the yield in those 20 cycles will be only 1.8520, or 220,513 copies of the starting material. In other words, a PCR running at 85 % efficiency will yield only 21 % as much final product, compared to a reaction running at 100 % efficiency. A reaction that is reduced to 50 % mean efficiency will yield less than 1 % of the possible product.
In practice, routine polymerase chain reactions rarely achieve the theoretical maximum yield, and PCRs are usually run for more than 20 cycles to compensate for the lower yield. At 50 % mean efficiency, it would take 34 cycles to achieve the milliorrfold amplification theoretically possible in 20, and at lower efficiencies, the number of cycles required becomes prohibitive. In addition, any background products that amplify with a better mean efficiency than the intended target will become the dominant products.
Also, many variables can influence the mean efficiency of PCR, including target DNA
length and secondary structure, primer length and design, primer and dNTP
concentrations, and buffer composition, to name but a few. Contamination of the reaction with exogenous DNA
(e.g., DNA spilled onto lab surfaces) or cross-contamination is also a major consideration.
Reaction conditions must be carefully optimized for each different primer pair and target sequence, and the process can take days, even for an experienced investigator.
The laboriousness of this process, including numerous technical considerations and other factors, presents a significant drawback to using PCR in the clinical setting. Indeed, PCR has yet to penetrate the clinical market in a significant way. The same concerns arise with LCR, as LCR
must also be optimized to use different oligonucleotide sequences for each target sequence. In addition, both methods require expensive equipment, capable of precise temperature cycling.
Many applications of nucleic acid detection technologies, such as in studies of allelic variation, involve not only detection of a specific sequence in a complex background, but also the discrimination between sequences with few, or single, nucleotide differences. One method of the detection of allele-specific variants by PCR is based upon the fact that it is difficult for Taq polymerase to synthesize a DNA strand when there is a mismatch between the template strand and the 3' end of the primer. An allele-specific variant may be detected by the use of a primer that is perfectly matched with only one of the possible alleles; the mismatch to the other allele acts to prevent the extension of the primer, thereby preventing the amplification of that sequence.
This method has a substantial limitation in that the base composition of the mismatch influences the ability to prevent extension across the mismatch, and certain mismatches do not prevent extension or have only a minimal effect.
A similar 3'-mismatch strategy is used with greater effect to prevent ligation in the LCR.
Any mismatch effectively blocks the action of the thermostable ligase, but LCR
still has the drawback of target-independent background ligation products initiating the amplification.
Moreover, the combination of PCR with subsequent LCR to identify the nucleotides at individual positions is also a clearly cumbersome proposition for the clinical laboratory.
The direct detection method according to various preferred embodiments of the present invention may be, for example a cycling probe reaction (CPR) or a branched DNA
analysis.
When a sufficient amount of a nucleic acid to be detected is available, there are advantages to detecting that sequence directly, instead of making more copies of that target, (e.g., as in PCR and LCR). Most notably, a method that does not amplify the signal exponentially is more amenable to quantitative analysis. Even if the signal is enhanced by attaching multiple dyes to a single oligonucleotide, fie correlation between the final signal intensity and amount of target is direct. Such a system has an additional advantage that the products of the reaction will not themselves promote further reaction, so contamination of lab 1 S surfaces by the products is not as much of a concern. Recently devised techniques have sought to eliminate the use of radioactivity and/or improve the sensitivity in automatable formats. Two examples are the "Cycling Probe Reaction" (CPR), and "Branched DNA" (bDNA).
Cycling probe reaction (CPR): The cycling probe reaction (CPR), uses a long chimeric oligonucleotide in which a central portion is made of RNA while the two termini are made of DNA. Hybridization of the probe to a target DNA and exposure to a thermostable RNase H
causes the RNA portion to be digested. 'This destabilizes the remaining DNA
portions of the duplex, releasing the remainder of the probe from the target DNA and allowing another probe molecule to repeat the process. The signal, in the form of cleaved probe molecules, accumulates at a linear rate. While the repeating process increases the signal, the RNA
portion of the oligonucleotide is vulnerable to RNases that may carried through sample preparation.
Branched DNA: Branched DNA (bDNA), involves oligonucleotides with branched structures that allow each individual oligonucleotide to carry 35 to 40 labels (e.g., alkaline phosphatase enzymes). While this enhances the signal from a hybridization event, signal from nonspecific binding is similarly increased.
The detection of at least one sequence -change according to various preferred embodiments of the present invention may be accomplished by, for example restriction fragment length polymorphism (RFLP analysis), allele specific oligonucleotide (ASO) analysis, Denaturing/Temperature Gradient Gel Electrophoresis (DGGE/TGGE), Single-Strand Conformation Polymorphism (SSCP) analysis or Dideoxy fingerprinting (ddF).
The demand for tests which allow the detection of specific nucleic acid sequences and sequence changes is growing rapidly in clinical diagnostics. As nucleic acid sequence data for genes from humans and pathogenic organisms accumulates, the demand for fast, cost-effective, and easy to-use tests for as yet mutations within specific sequences is rapidly increasing.
A handful of methods have been devised to scan nucleic acid segments for mutations.
One option is to determine the entire gene sequence of each test sample (e.g., a bacterial isolate).
For sequences under approximately 600 nucleotides, this may be accomplished using amplified material (e.g., PCR reaction products). This avoids the time and expense associated with cloning the segment of interest. However, specialized equipment and highly trained personnel are required, and the method is too labor-intense and expensive to be practical and effective in the clinical setting.
In view of the difficulties associated with sequencing, a given segment of nucleic acid may be characterized on several other levels. At the lowest resolution, the size of the molecule can be determined by electrophoresis by comparison to a known standard run on the same gel. A
more detailed picture of the molecule may be achieved by cleavage with combinations of restriction enzymes prior to electrophoresis, to allow construction of an ordered map. The presence of specific sequences within the fragment can be detected by hybridization of a labeled probe, or the precise nucleotide sequence can be determined by partial chemical degradation or by primer extension in the presence of chain-terminating nucleotide analogs.
Restriction fragment length polymorphism (RFLP): For detection of single-base differences between like sequences, the requirements of the analysis are often at the highest level of resolution. For cases in which the position of the nucleotide in question is known in advance, several methods have been developed for examining single base changes without direct sequencing. For example, if a mutation of interest happens to fall within a restriction recognition sequence, a change in the pattern of digestion can be used as a diagnostic tool (e.g., restriction fragment length polymorphism [RFLP] analysis).
Single point mutations have been also detected by the creation or destruction of RFLPs.
Mutations are detected and bcalized by the presence and size of the RNA
fragments generated by cleavage at the mismatches. Single nucleotide mismatches in DNA
heteroduplexes are also recognized and cleaved by some chemicals, providing an alternative strategy to detect single base substitutions, generically named the "Mismatch Chemical Cleavage" (MCC).
However, this method requires the use of osmium tetroxide and piperidine, two highly noxious chemicals which are not suited for use in a clinical laboratory.
RFLP analysis suffers from low sensitivity and requires a large amount of sample. When RFLP analysis is used for the detection of point mutations, it is, by its nature, limited to the detection of only those single base changes which fall within a restriction sequence of a known restriction endonuclease. Moreover, the majority of the available enzymes have 4 to 6 base-pair recognition sequences, and cleave too frequently for many large-scale DNA
manipulations.
Thus, it is applicable only in a small fraction of cases, as most mutations do not fall within such sites.
A handful of rare-cutting restriction enzymes with 8 base-pair specificities have been isolated and these are widely used in genetic mapping, but these enzymes are few in number, are I S limited to the recognition of G+C-rich sequences, and cleave at sites that tend to be highly clustered. Recently, endonucleases encoded by group I introns have been discovered that might have greater than 12 base-pair specificity, but again, these are few in number.
Allele specific oligonucleotide (ASO): If the change is not in a recognition sequence, then allele-specific oligonucleotides (ASOs), can be designed to hybridize in proximity to the mutated nucleotide, such that a primer extension or ligation event can bused as the indicator of a match or a mis-match. Hybridization with radioactively labeled allelic specific oligonucleotides (ASO) also has been applied to the detection of specific point mutations. The method is based on the differences in the melting temperature of short DNA fragments differing by a single nucleotide. Stringent hybridization and washing conditions can differentiate between mutant and wild-type alleles. The ASO approach applied to PCR products also has been extensively utilized by various researchers to detect and characterize point mutations in ras genes and gsp/gip oncogenes. Because of the presence of various nucleotide changes in multiple positions, the ASO method requires the use of many oligonucleotides to cover all possible oncogenic mutations.
With either of the techniques described above (i.e., RFLP and ASO), the precise location of the suspected mutation must be known in advance of the test. That is to say, they are inapplicable when one needs to detect the presence of a mutation within a gene or sequence of interest.
DenaturinglTemperature Gradient Gel Electrophoresis (DGGElTGGE): Two other methods rely on detecting changes in electrophoretic mobility in response to minor sequence changes. One of these methods, termed "Denaturing Gradient Gel Electrophoresis" (DGGE) is based on the observation that slightly different sequences will display different patterns of local melting when electrophoretically resolved on a gradient gel. In this manner, variants can be distinguished, as differences in melting properties of homoduplexes versus heteroduplexes differing in a single nucleotide can detect the presence of mutations in the target sequences because of the corresponding changes in their electrophoretic mobilities. The fragments to be analyzed, usually PCR products, are "clamped" at one end by a long stretch of CtC base pairs (30-80) to allow complete denaturation of the sequence of interest without complete dissociation of the strands. The attachment of a GC "clamp" to the DNA fragments increases the faction of mutations that can be recognized by DGGE. Attaching a GC clamp to one primer is critical to ensure that the amplified sequence has a low dissociation temperature.
Modifications of the technique have been developed, using temperature gradients, and the method can be also applied to RNA:RNA duplexes.
Limitations on the utility of DGGE include the requirement that the denaturing conditions must be optimized for each type of DNA to be tested. Furthermore, the method requires specialized equipment b prepare the gels and maintain the needed high temperatures during electrophoresis. The expense associated with the synthesis of the clamping tail on one oligonucleotide for each sequence to be tested is also a major consideration.
In addition, long running times are required for DGGE. The long running time of DGGE was shortened in a modification of DGGE called constant denaturant gel electrophoresis (CDGE).
CDGE requires that gels be performed under different denaturant conditions in order to reach high efficiency for the detection of mutations.
A technique analogous to DGGE, termed temperature gradient gel electrophoresis (TGGE), uses a thermal gradient rather than a chemical denaturant gradient.
TGGE requires the use of specialized equipment which can generate a temperature gradient perpendicularly oriented relative to the electrical field. TGGE can detect mutations in relatively small fragments of DNA
therefore scanning of large gene segments requires the use of multiple PCR
products prior to running the gel.
Single-Strand Conformation Polymorphism (SSCP): Another common method, called "Single-Strand Conformation Polymorphism" (SSCP) was developed by Hayashi, Sekya and colleagues and is based on the observation that single strands of nucleic acid can take on characteristic conformations in non-denaturing conditions, and these conformations influence electrophoretic mobility. The complementary strands assume sufficiently different structures that one strand may be resolved from the other. Changes in sequences within the fragment will also change the conformation, consequently altering the mobility and allowing this to be used as an assay for sequence variations.
The SSCP process involves denaturing a DNA segment (e.g., a PCR product) that is labeled on both strands, followed by slow electrophoretic separation on a norrdenaturing polyacrylamide gel, so that intra-molecular interactions can form and not be disturbed during the run. This technique is extremely sensitive to variations in gel composition and temperature. A
serious limitation of this method is the relative difficulty encountered in comparing data generated in different laboratories, under apparently similar conditions.
Dideoxy fingerprinting (ddF): The dideoxy fingerprinting (ddF) is another technique developed to scan genes for the presence of mutations. The ddF technique combines components of Sanger dideoxy sequencing with SSCP. A dideoxy sequencing reaction is performed using one dideoxy terminator and then the reaction products are electrophoresed on nondenaturing polyacrylamide gels to detect alterations in mobility of the termination segments as in SSCP analysis. While ddF is an improvement over SSCP in terms of increased sensitivity, ddF requires the use of expensive dideoxynucleotides and this technique is still limited to the analysis of fragments of the size suitable for SSCP (i.e., fragments of 200-300 bases for optimal detection of mutations).
In addition to the above limitations, all of these methods are limited as to the size of the nucleic acid fragment that can be analyzed. For the direct sequencing approach, sequences of greater than 600 base pairs require cloning, with the consequent delays and expense of either deletion sub-cloning or primer walking, in order b cover the entire fragment.
SSCP and DGGE
have even more severe size limitations. Because of reduced sensitivity to sequence changes, these methods are not considered suitable for larger fragments. Although SSCP
is reportedly able to detect 90 % of single-base substitutions within a 200 base-pair fragment, the detection drops to less than 50 % for 400 base pair fragments. Similarly, the sensitivity of DGGE decreases as the length of the fragment reaches 500 base-pairs. The ddF technique, as a combination of direct sequencing and SSCP, is also limited by the relatively small size of the DNA
that can be screened.
According to a presently preferred embodiment of the present invention the step of searching for any of the nucleic acid sequences described here, in tumor cells or in cells derived from a cancer patient is effected by any suitable technique, including, but not limited to, nucleic acid sequencing, polymerise chain reaction, ligase chain reaction, self sustained synthetic reaction, Q(3-Replicase, cycling probe reaction, branched DNA, restriction fragment length polymorphism analysis, mismatch chemical cleavage, heteroduplex analysis, allele-specific oligonucleotides, denaturing gradient gel electrophoresis, constant denaturant gel electrophoresis, temperature gradient gel electrophoresis and dideoxy fingerprinting.
Detection may also optionally be performed with a chip or other such device.
The nucleic acid sample which includes the candidate region to be analyzed is preferably isolated, amplified and labeled with a reporter group. This reporter group can be a fluorescent group such as phycoerythrin. The labeled nucleic acid is then incubated with the probes immobilized on the chip using a fluidics station. describe the fabrication of fluidics devices and particularly microcapillary devices, in silicon and glass substrates.
Once the reaction is completed, the chip is inserted into a scanner and patterns of hybridization are detected. The hybridization data is collected, as a signal emitted from the reporter groups already incorporated into the nucleic acid, which is now bound to the probes attached to the chip. Since the sequence and position of each probe immobilized on the chip is known, the identity of the nucleic acid hybridized to a given probe can be determined.
It will be appreciated that when utilized along with automated equipment, the above described detection methods can be used to screen multiple samples for a disease and/or pathological condition both rapidly and easily.
Amino acid sequences and peptides The terms "polypeptide," "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an analog or mimetic of a con-esponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers.
Polypeptides can be modified, e.g., by the addition of carbohydrate residues to form glycoproteins. The terms "polypeptide," "peptide" and "protein" include glycoproteins, as well as non-glycoproteins.
Polypeptide products can be biochemically synthesized such as by employing standard solid phase techniques. Such methods include but are not limited to exclusive solid phase synthesis, partial solid phase synthesis methods, fragment condensation, classical solution synthesis. These methods are preferably used when the peptide is relatively short (i.e., 10 kDa) and/or when it cannot be produced by recombinant techniques (i.e., not encoded by a nucleic acid sequence) and therefore involves different chemistry.
Solid phase polypeptide synthesis procedures are well known in the art and further described by John Morrow Stewart and Janis Dillaha Young, Solid Phase Peptide Syntheses (2nd Ed., Pierce Chemical Company, 1984).
Synthetic polypeptides can optionally be purified by preparative high performance liquid chromatography [Creighton T. (1983) Proteins, structures arid molecular principles. WH
Freeman and Co. N.Y.], after which their composition can be confirmed via amino acid sequencing.
In cases where large amounts of a polypeptide are desired, it can be generated using recombinant techniques such as described by Bitter et al., (1987) Methods in Enzymol. 153:516-544, Studier et al. (1990) Methods in Enzymol. 185:60-89, Brisson et al.
(1984) Nature 310:511-514, Takamatsu et al. (1987) EMBO J. 6:307-311, Coruzzi et al. (1984) EMBO J.
3:1671-1680 and Brogli et al., (1984) Science 224:838-843, Gurley et al. (1986) Mol. Cell.
Biol. 6:559-565 and Weissbach & Weissbach, 1988, Methods for Plant Molecular Biology, Academic Press, NY, Section VIII, pp 421-463.
The present invention also encompasses polypeptides encoded by the polynucleotide sequences of the present invention, as well as polypeptides according to the amino acid sequences described herein. The present invention also encompasses homologues of these polypeptides, such homologues can be at least 50 %, at least 55 %, at least 60%, at least 65 %, at least 70 %, at least 75 %, at least 80 %, at least 85 %, at least 95 % or more say 100 homologous to the amino acid sequences set forth below, as can be determined using BlastP
software of the National Center of Biotechnology Information (NCBI) using default parameters, optionally and preferably including the following: filtering on (this option filters repetitive or low-complexity sequences from the query using the Seg (protein) program), scoring matrix is BLOSUM62 for proteins, word size is 3, E value is 10, gap costs are I1, 1 (initialization and extension), and number of alignments shown is 50. Nucleotide (nucleic acid) sequence S homology/identity is preferably determined by using the BlastN software of the National Center of Biotechnology Information (NCBI) using default parameters, which preferably include using the DUST filter program, and also preferably include having an E value of 10, filtering low complexity sequences and a word size of 11. Finally, the present invention also encompasses fragments of the above described polypeptides and polypeptides having mutatio ns, such as deletions, insertions or substitutions of one or more amino acids, either naturally occurring or artificially induced, either randomly or in a targeted fashion.
It will be appreciated that peptides identified according the present invention may be degradation products, synthetic peptides or recombinant peptides as well as peptidomimetics, typically, synthetic peptides and peptoids and semipeptoids which are peptide analogs, which I S may have, for example, modifications rendering the peptides more fable while in a body or more capable of penetrating into cells. Such modifications include, but are not limited to N
terminus modification, C terminus modification, peptide bond modification, including, but not limited to, CH2-NH, CH2-S, CH2-S=O, O=C-NH, CH2-O, CH2-CH2, S=C-NH, CH=CH or CF=CH, backbone modifications, and residue modification. Methods for preparing peptidomimetic compounds are well known in the art and are specified. Further details in this respect are provided hereinunder.
Peptide bonds (-CO-NH-) within the peptide may be substituted, for example, by I~
methylated bonds ( N(CH3)-CO-), ester bonds ( C(R)H-C-O-O-C(R)-N-), ketomethylen bonds (-CO-CH2-), oc-aza bonds (-NH-N(R)-CO-), wherein R is any alkyl, e.g, methyl, carba bonds CH2-NH-), hydroxyethylene bonds (-CH(OH)-CH2-), thioamide bonds (-CS-NH-), olefinic double bonds (-CH=CH-), retro amide bonds (-NH-CO-), peptide derivatives (-N(R)-CH2-CO-), wherein R is the "normal" side chain, naturally presented on the carbon atom.
These modifications can occur at any of the bonds along the peptide chain and even at several (2-3) at the same time.
Natural aromatic amino acids, Trp, Tyr and Phe, may be substituted for synthetic norr natural acid such as Phenylglycine, TIC, naphthylelanine (Nol), ring-methylated derivatives of Phe, halogenated derivatives of Phe or o-methyl-Tyr.
In addition to the above, the peptides of the present invention may also include one or more modified amino acids or one or more norramino acid monomers (e.g. fatty acids, complex carbohydrates etc).
As used herein in the specification and in the claims section below the term "amino acid"
or "amino acids" is understood to include the 20 naturally occurring amino acids; those amino acids often modified post-translationally in vivo, including, for example, hydroxyproline, phosphoserine and phosphothreonine; and other unusual amino acids including, but not limited to, 2-aminoadipic acid, hydroxylysine, isodesmosine, nor-valine, nor-leucine and ornithine.
Furthermore, the term "amino acid" includes both D- and Lramino acids.
Table I non-conventional or modified amino acids which can be used with the present invention.
Table 1 Non-conventional Code Non-conventional Code amino amino acid acid a-aminobutyric Abu LrN-methylalanine Nmala acid a-amino-a-methylbutyrateMgabu >~N-methylarginine Nmarg aminocyclopropane-Cpro IrN-methylasparagineNmasn Carboxylate LrN-methylaspartic Nmasp acid aminoisobutyric Aib LrN-methylcysteine Nmcys acid aminonorbornyl- Norb LrN-methylglutamine Nmgin Carboxylate LrN-methylglutamic Nmglu acid Cyclohexylalanine Chexa IrN-methylhistidine Nmhis CyclopentylalanineCpen LrN-methylisolleucineNmile D-alanine Dal LrN-methylleucine Nmleu D-arginine Darg LrN-methyllysine Nmlys D-aspartic acid Dasp L-N-methylmethionineNmmet D-cysteine Dcys L-N-methylnorleucineNmnle D-glutamine Dgln L-N-methylnorvalineNmova D-glutamic acid Dglu L-N-methylornithineNmorn D-histidine Dhis L-N-methylphenylalanineNmphe D-isoleucine Dile L-N-methylproline Nmpro D-leucine Dleu L-N-methylserine Nmser D-lysine Dlys L-N-methylthreonineNmthr D-methionine Dmet L-N-methyltryptophanNmtrp D-ornithine Dorn L-N-methyltyrosine Nmtyr D-phenylalanine Dphe L-N-methylvaline Nmval D-proline Dpro L-N-methylethylglycineNmetg D-serine Dser IrN-methyl-t-butylglycineNmtbug D-threonine Dthr Lrnorleucine N]e D-tryptophan Dtrp Lrnorvaline Nva D-tyrosine Dtyr a-methyl-aminoisobutyrateMaib D-valine Dval a-methyl-y-aminobutyrateMgabu D-a-methylalanine Dmala a-methylcyclohexylalanineMchexa D-a-methylarginineDmarg a-methylcyclopentylalanineMcpen D-a-methylasparagineDmasn a-methyl-a-napthylalanineManap D-a-methylaspartateDmasp a- methylpenicillamineMpen D-a-methylcysteineDmcys N-(4-aminobutyl)glycineNglu D-a-methylglutamineDmgln N-(2-aminoethyl)glycineNaeg D-a-methylhistidineDmhis N-(3-aminopropyl)glycineNorn D-a-methylisoleucineDmile N- amino-a-methylbutyrateNmaabu D-a-methylleucine Dmleu a-napthylalanine Anap D-a-methyllysine Dmlys N-benzylglycine Nphe D-a-methylmethionineDmmet N-(2-carbamylethyl)glycineNgln D-a-methylornithineDmorn N-(carbamylmethyl)glycineNasn D-a-methylphenylalanineDmphe N-(2-carboxyethyl)glycineNglu D-a-methylprolineDmpro N-(carboxymethyl)glycineNasp D-a-methylserine Dmser N-cyclobutylglycineNcbut D-a-methylthreonineDmthr N-cycloheptylglycineNchep D-a-methyltryptophanDmtrp N-cyclohexylglycineNchex D-a-methyltyrosineDmty N-cyclodecylglycineNedec D-a-methylvaline Dmval N-cyclododeclglycineNcdod D-a-methylalnine Dnmala N-cyclooctylglycineNcoct D-a-methylarginineDnmarg N-cyclopropylglycineNcpro D-a-methylasparagineDnmasn N-cycloundecylglycineNcund D-a-methylasparatateDnmasp N-(2,2-diphenylethyl)glycineNbhm D-a-methylcysteineDnmcys N-(3,3- Nbhe diphenylpropyl)glycine D-N-methylleucineDnmleu N-(3-indolylyethyl)Nhtrp glycine D-N-methyllysine Dnmlys N-methyl-y-aminobutyrateNmgabu N- Nmchexa D-N-methylmethionineDnmmet methylcyclohexylalanine D-N-methylornithineDnmorn N-methylcyclopentylalanineNmcpen N-methylglycine Nala D-N-methylphenylalanineDnmphe N-methylaminoisobutyrateNmaib D-N-methylproline Dnmpro N-(1-methylpropyl)glycineNile D-N-methylserine Dnmser N-(2-methylpropyl)glycineNile D-N-methylserine Dnmser N-(2-methylpropyl)glycineNleu D-N-methylthreonineDnmthr D-N-methyltryptophanDnmtrp N-(1-methylethyl)glycineNva D-N-methyltyrosineDnmtyr N-methyla-napthylalanineNmanap D-N-methylvaline Dnmval N-methylpenicillamineNmpen y-aminobutyric Gabu N-(p-hydroxyphenyl)glycineNhtyr acid Lrt-butylglycine Tbug N-(thiomethyl)glycineNcys Irethylglycine Etg penicillamine Pen L-homophenylalanineHphe L-a-methylalanine Mala L-a-methylarginineMarg L-a-methylasparagineMasn L-a-methylaspartateMasp L-a-methyl-t-butylglycineMtbug L-a-methylcysteineMcys L-methylethylglycineMetg L-a-methylglutamineMgln L-a-methylglutamateMglu L-a-methylhistidineMhis L-a-methylhomo Mhphe phenylalanine L-a-methylisoleucineMile N-(2-methylthioethyl)glycineNmet D-N-methylglutamineDnmgln N-(3- Narg guanidinopropyl)glycine D-N-methylglutamateDnmglu N-(1-hydroxyethyl)glycineNthr D-N-methylhistidineDnmhis N-(hydroxyethyl)glycineNser D-N-methylisoleucineDnmile N-(imidazolylethyl)glycineNhis D-N-methylleucine Dnmleu N-(3-indolylyethyl)glycineNhtrp D-N-methyllysine Dnmlys N-methyl-'y-aminobutyrateNmgabu N- Nmchexa D-N-methylmethionineDnmmet methylcyclohexylalanine D-N-methylornithineDnmorn N-methylcyclopentylalanineNmcpen N-methylglycine Nala D-N-methylphenylalanineDnmphe N-methylaminoisobutyrateNmaib D-N-methylproline Dnmpro N-(1-methylpropyl)glycineNile D-N-methylserine Dnmser N-(2-methylpropyl)glycineNleu D-N-methylthreonineDnmthr D-N-methyltryptophanDnmtrp N-(1-methylethyl)glycineNval D-N-methyltyrosineDnmtyr N-methyla-napthylalanineNmanap D-N-methylvaline Dnmval N-methylpenicillamineNmpen 'y-aminobutyric Gabu N-(p-hydroxyphenyl)glycineNhtyr acid Lrt-butylglycine Tbug N-(thiomethyl)glycineNcys L-ethylglycine Etg penicillamine Pen IrhomophenylalanineHphe Ira-methylalanine Mala L-a-methylarginineMarg L-a-methylasparagineMasn L-a-methylaspartateMasp L-a-methyl-t-butylglycineMtbug L-a-methylcysteineMcys L-methylethylglycineMetg L-a-methylglutamineMgln L-a-methylglutamateMglu L-a-methylhistidineMhis L-a- Mhphe methylhomophenylalanine L-a-methylisoleucineMile N-(2-methylthioethyl)glycineNmet L-a-methylleucineMleu Lra-methyllysine Mlys L-a-methylmethionineMmet L-a-methylnorleucineMnle L-a-methylnorvalineMnva L-a-methylornithineMorn L-a-methylphenylalanineMphe L-a-methylproline Mpro L-a-methylserine mser L-a-methylthreonineMthr Ira-methylvaline Mtrp L-a-methyltyrosine Mtyr Lra-methylleucineMval L-N- Nmhphe Nnbhm methylhomophenylalanine N-(N-(2,2-diphenylethyl) N-(N-(3,3-diphenylpropyl) carbamylmethyl-glycineNnbhm carbamylmethyl(1)glyeineNnbhe 1-carboxy-1-(2,2-diphenylNmbc ethylamino)cyclopropane Table 1 Cont.
Since the peptides of the present invention are preferably utilized in diagnostics which require the peptides to be in soluble form, the peptides of the present invention preferably include one or more norrnatural or natural polar amino acids, including but not limited to serine and threonine which are capable of increasing peptide solubility due to their hydroxyl-containing side chain.
The peptides of the present invention are preferably utilized in a linear form, although it will be appreciated that in cases where cyclicization does not severely interfere with peptide characteristics, cyclic forms of the peptide can also be utilized.
The peptides of present invention can be biochemically synthesized such as by using standard solid phase techniques. These methods include exclusive solid phase synthesis well known in the art, partial solid phase synthesis methods, fragment condensation, classical solution synthesis. These methods are preferably used when the peptide is relatively short (i.e., 10 kDa) and/or when it cannot be produced by recombinant techniques (i.e., not encoded by a nucleic acid sequence) and therefore involves different chemistry.
Synthetic peptides can be purified by preparative high performance liquid chromatography and the composition of which can be confirmed via amino acid sequencing.
In cases where large amounts of the peptides of the present invention are desired, the peptides of the present invention can be generated using recombinant techniques such as described by Bitter et al., (1987) Methods in Enzymol. 153:516-544, Studier et al. (1990) Methods in Enzymol. 185:60-89, Brisson et al. (1984) Nature 310:511-514, Takamatsu et al.
(1987) EMBO J. 6:307-311, Coruzzi et al. (1984) EMBO J. 3:1671-1680 and Brogli et al., (1984) Science 224:838-843, Gurley et al. (1986) Mol. Cell. Biol. 6:559-565 and Weissbach &
Weissbach, 1988, Methods for Plant Molecular Biology, Academic Press, NY, Section VIII, pp 421-463 and also as described above.
Antibodies "Antibody" refers to a polypeptide ligand that is preferably substantially encoded by an immunoglobulin gene or immunoglobulin genes, or fragments thereof, which specifically binds and recognizes an epitope (e.g., an antigen). The recognized immunoglobulin genes include the kappa and lambda light chain constant region genes, the alpha, gamma, delta, epsilon and mu heavy chain constant region genes, and the myriad-immunoglobulin variable region genes.
Antibodies exist, e.g., as intact immunoglobulins or as a number of well characterized fragments produced by digestion with various peptidases. This includes, e.g., Fab' and F(ab)'z fragments.
The term "antibody," as used herein, also includes antibody fragments either produced by the modification of whole antibodies or those synthesized de novo using recombinant DNA
methodologies. It also includes polyclonal antibodies, monoclonal antibodies, chimeric antibodies, humanized antibodies, or single chain antibodies. "Fc" portion of an antibody refers to that portion of an immunoglobulin heavy chain that comprises one or more heavy chain constant region domains, CHI, CH2 and CH3, but does not include the heavy chain variable region.
The functional fragments of antibodies, such as Fab, F(ab')2, and Fv that are capable of binding to macrophages, are described as follows: (1) Fab, the fragment which contains a monovalent antigen-binding fragment of an antibody molecule, can be produced by digestion of whole antibody with the enzyme papain to yield an intact light chain and a portion of one heavy chain; (2) Fab', the, fragment of an antibody molecule that can be obtained by treating whole antibody with pepsin, followed by reduction, to yield an intact light chain and a portion of the heavy chain; two Fab' fragments are obtained per antibody molecule; (3) (Fab')2, the fragment of the antibody that can be obtained by treating whole antibody with the enzyme pepsin without subsequent reduction; F(ab')2 is a dimer of two Fab' fragments held together by two disulfide bonds; (4) Fv, defined as a genetically engineered fragment containing the variable region of the light chain and the variable region of the heavy chain expressed as two chains; and (5) Single chain antibody ("SCA"), a genetically engineered molecule containing the variable region of the light chain and the variable region of the heavy chain, linked by a suitable polypeptide linker as a genetically fused single chain molecule.
Methods of producing polyclonal and monoclonal antibodies as well as fragments thereof are well known in the art (See for example, Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York, 1988, incorporated herein by reference).
Antibody fragments according to the present invention can be prepared by proteolytic hydrolysis of the antibody or by expression in E. coli or mammalian cells (e.g. Chinese hamster ovary cell culture or other protein expression systems) of DNA encoding the fragment.
Antibody fragments can be obtained by pepsin or papain digestion of whole antibodies by conventional methods. For example, antibody fragments can be produced by enzymatic cleavage of antibodies with pepsin to provide a SS fragment denoted F(ab')2.
This fragment can be further cleaved using a thiol reducing agent, and optionally a blocking group for the sulfhydryl groups resulting from cleavage of disulfide linkages, to produce 3.SS Fab' monovalent fragments. Alternatively, an enzymatic cleavage using pepsin produces two monovalent Fab' fragments and an Fc fragment directly. These methods are described, for example, by Goldenberg, U.S. Pat. Nos. 4,036,945 and 4,331,647, and references contained therein, which patents are hereby incorporated by reference in their entirety.
See also Porter, R.
R. [Biochem. J. 73: 1 19-126 ( 1959)]. Other methods of cleaving antibodies, such as separation of heavy chains to form monovalent light-heavy chain fragments, further cleavage of fragments, or other enzymatic, chemical, or genetic techniques may also be used, so long as the fragments bind to the antigen that is recognized by the intact antibody.
Fv fragments comprise an association of VH and VL chains. This association may be noncovalent, as described in hbar et al. [Proc. Nat'1 Acad. Sci. USA 69:2659-62 (19720].
Alternatively, the variable chains can be linked by an intermolecular disulfide bond or cross-linked by chemicals such as glutaraldehyde. Preferably, the Fv fragments comprise VH and VL
chains connected by a peptide linker. These single-chain antigen binding proteins (sFv) are prepared by constructing a structural gene comprising DNA sequences encoding the VH and VL
domains connected by an oligonucleotide. The structural gene is inserted into an expression vector, which is subsequently introduced into a host cell such as E. coli. The recombinant host cells synthesize a single polypeptide chain with a linker peptide bridging the two V domains.
Methods for producing sFvs are described, for example, by [Whitlow and Filpula, Methods 2:
97-105 (1991); Bird et al., Science 242:423-426 (1988); Pack et al., Bio/Technology 11:1271-77 (1993); and U.S. Pat. No. 4,946,778, which is hereby incorporated by reference in its entirety.
Another form of an antibody fragment is a peptide coding for a single complementarit~
determining region (CDR). CDR peptides ("minimal recognition units") can be obtained by constructing genes encoding the CDR of an antibody of interest. Such genes are prepared, for example, by using the polymerise chain reaction to synthesize the variable region from RNA of antibody producing cells. See, for example, Larrick and Fry [Methods, 2: 106-10 (1991)].
Humanized forms of non-human (e.g., murine) antibodies are chimeric molecules of immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab') or other antigen-binding subsequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin. Humanized antibodies include human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a norrhuman species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity. In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding norrhuman residues. Humanized antibodies may also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin [Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol., 2:593-596 (1992)].
Methods for humanizing norrhuman antibodies are well known in the art.
Generally, a humanized antibody has one or more amino acid residues introduced into it from a source which is norrhuman. These non-human amino acid residues are often referred to as import residues, which are typically taken from an import variable domain. Humanization can be essentially performed following the method of Winter and co-workers [Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)], by substituting rodent CDRs or CDR sequences for the corresponding sequences IS of a human antibody. Accordingly, such humanized antibodies are chimeric antibodies (U.S.
Pat. No. 4,816,567), wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR
residues are substituted by residues from analogous sites in rodent antibodies.
Human antibodies can also be produced using various techniques known in the art, including phage display libraries [Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991);
Marks et al., J. Mol. Biol., 222:581 (1991)]. The techniques of Cole et al.
and Boerner et al. are also available for the preparation of human monoclonal antibodies (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985) and Boerner et al., J. Immunol., 147(1):86-95 (1991)]. Similarly, human antibodies can be made by introduction of human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126;
5,633,425;
5,661,016, and in the following scientific publications: Marks et al., Bio/Technology 10,: 779-783 ( I 992); Lonberg et al., Nature 368: 856-859 ( 1994); Morrison, Nature 368 812- I 3 ( 1994);
Fishwild et al., Nature Biotechnology 14, 845-51 (1996); Neuberger, Nature Biotechnology 14:
826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13, 65-93 (1995).
Preferably, the antibody of this aspect of the present invention specifically binds at least one epitope of the polypeptide variants of the present invention. As used herein, the term "epitope" refers to any antigenic determinant on an antigen to which the paratope of an antibody binds.
Epitopic determinants usually consist of chemically active surface groupings of molecules such as amino acids or carbohydrate side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics.
Optionally, a unique epitope may be created in a variant due to a change in one or more post-translational modifications, including but not limited to glycosylation and/or phosphorylation, as described below. Such a change may also cause a new epitope to be created, for example through removal of glycosylation at a particular site.
An epitope according to the present invention may also optionally comprise part or all of a unique sequence portion of a variant according to the present invention in combination with at least one other portion of the variant which is not contiguous to the unique sequence portion in the linear polypeptide itself, yet which are able to form an epitope in combination. One or more unique sequence portions may optionally combine with one or more other noncontiguous portions of the variant (including a portion which may have high homology to a portion of the known protein) to form an epitope.
Immunoassays In another embodiment of the present invention, an immunoassay can be used to qualitatively or quantitatively detect and analyze markers in a sample. This method comprises:
providing an antibody that specifically binds to a marker; contacting a sample with the antibody;
and detecting the presence of a complex of the antibody bound to the marker in the sample.
To prepare an antibody that specifically binds to a marker, purified protein markers can be used. Antibodies that specifically bind to a protein marker can be prepared using any suitable methods known in the art.
After the antibody is provided, a marker can be detected and/or quantified using any of a number of well recognized immunological binding assays. Useful assays include, for example, an enzyme immune assay (EIA) such as enzyme-linked immunosorbent assay (ELISA), a radioimmune assay (RIA), a Western blot assay, or a slot blot assay see, e.g., U.S. Pat. Nos.
4,366,241; 4,376,110; 4,517,288; and 4,837,168). Generally, a sample obtained from a subject can be contacted with the antibody that specifically binds the marker.
Optionally, the antibody can be fixed to a solid support to facilitate washing and subsequent isolation of the complex, prior to contacting the antibody with a sample. Examples of solid supports include but are not limited to glass or plastic in the form of, e.g., a microtiter plate, a stick, a bead, or a microbead. Antibodies can also be attached to a solid support.
After incubating the sample with antibodies, the mixture is washed and the antibody marker complex formed can be detected. This can be accomplished by incubating the washed mixture with a detection reagent. Alternatively, the marker in the sample can be detected using an indirect assay, wherein, for example, a second, labeled antibody is used to detect bound marker-specific antibody, and/or in a competition or inhibition assay wherein, for example, a monoclonal antibody which binds to a distinct epitope of the marker are incubated simultaneously with the mixture.
Throughout the assays, incubation and/or washing steps may be required after each combination of reagents. Incubation steps can vary from about 5 seconds to several hours, preferably from about 5 minutes to about 24 hours. However, the incubation time will depend upon the assay format, marker, volume of solution, concentrations and the like. Usually the assays will be earned out at ambient temperature, although they can be conducted over a range of temperatures, such as 10 °C to 40 °C.
The immunoassay can be used to determine a test amount of a marker in a sample from a subject. First, a test amount of a marker in a sample can be detected using the immunoassay methods described above. If a marker is present in the sample, it will form an antibody-marker complex with an antibody that specifically binds the marker under suitable incubation conditions described above. The amount of an antibody marker complex can optionally be determined by comparing to a standard. As noted above, the test amount of marker need not be measured in absolute units, as long as the unit of measurement can be compared to a control amount and/or signal.
Preferably used are antibodies which specifically interact with the polypeptides of the present invention and not with wild type proteins or other isoforms thereof, for example. Such antibodies are directed, for example, to the unique sequence portions of the polypeptide variants of the present invention, including but not limited to bridges, heads, tails and insertions described in greater detail below. Preferred embodiments of antibodies according to the present invention are described in greater detail with regard to the section entitled "Antibodies".
Radio-immunoassay (RIA): In one version, this method involves precipitation of the desired substrate and in the methods detailed hereinbelow, with a specific antibody and radiolabelled antibody binding protein (e.g., protein A labeled with 1125) immobilized on a precipitable earner such as agarose beads. The number of counts in the precipitated pellet is proportional to the amount of substrate.
In an alternate version of the RIA, a labeled substrate and an unlabelled antibody binding protein are employed. A sample containing an unknown amount of substrate is added in varying amounts. The decrease in precipitated counts from the labeled substrate 's proportional to the amount of substrate in the added sample.
Enzyme linked immunosorbent assay (ELISA): This method involves fixation of a sample (e.g., fixed cells or a proteinaceous solution) containing a protein substrate to a surface such as a well of a microtiter plate. A substrate specific antibody coupled to an enzyme is applied and allowed to bind to the substrate. Presence of the antibody is then detected and quantitated by a colorimetric reaction employing the enzyme coupled to the antibody. Enzymes commonly employed in this method include horseradish peroxidase and alkaline phosphatase. If well calibrated and within the linear range of response, the amount of substrate present in the sample is proportional to the amount of color produced. A substrate standard is generally employed to improve quantitative accuracy.
Western blot: This method involves separation of a substrate from other protein by means of an acrylamide gel followed by transfer of the substrate to a membrane (e.g., nylon or PVDF).
Presence of the substrate is then detected by antibodies specific to the substrate, which are in turn detected by antibody binding reagents. Antibody binding reagents may be, for example, protein A, or other antibodies. Antibody binding reagents may be radiolabelled or enzyme linked as described hereinabove. Detection may be by autoradiography, colorimetric reaction or chemiluminescence. This method allows both quantitation of an amount of substrate and determination of its identity by a relative position on the membrane which is indicative of a migration distance in the acrylamide gel during electrophoresis.
Irnmunohistochemical analysis: This method involves detection of a substrate in situ in fixed cells by substrate specific antibodies. The substrate specific antibodies may be enzyme linked or linked to fluorophores. Detection is by microscopy and subjective evaluation. If enzyme linked antibodies are employed, a colorimetric reaction may be required.
Fluorescence activated cell sorting (FRCS): This method involves detection of a substrate in situ in cells by substrate specific antibodies. The substrate specific antibodies are linked to fluorophores. Detection is by means of a cell sorting machine which reads the wavelength of light emitted from each cell as it passes through a light beam.
This method may employ two or more antibodies simultaneously.
1 S Radio-imaging Methods These methods include but are not limited to, positron emission tomography (PET) single photon emission computed bmography (SPELT). Both of these techniques are norr invasive, and can be used to detect and/or measure a wide variety of tissue events and/or functions, such as detecting cancerous cells for example. Unlike PET, SPELT
can optionally be used with two labels simultaneously. SPELT has some other advantages as well, for example with regard to cost and the types of labels that can be used. For example, US
Patent No.
6,696,686 describes the use of SPELT for detection of breast cancer, and is hereby incorporated by reference as if fully set forth herein.
Display Libraries According to still another aspect of the present invention there is provided a display library comprising a plurality of display vehicles (such as phages, viruses or bacteria) each displaying at least 6, at least 7, at least 8, at least 9, at least 10, 10-15, 12-17, 15-20, 15-30 or 20-50 consecutive amino acids derived from the polypeptide sequences of the present invention.
Methods of constructing such display libraries are well known in the art. Such methods are described in, for example, Young AC, et al., "The three-dimensional structures of a polysaccharide binding antibody to Cryptococcus neoformans and its complex with a peptide from a phage display library: implications for the identification of peptide mimotopes" J Mol Biol 1997 Dec 12;274(4):622-34; Giebel LB et al. "Screening of cyclic peptide phage libraries identifies ligands that bind streptavidin with high affinities" Biochemistry 1995 Nov 28;34(47):15430-5; Davies EL et al., "Selection of specific phage-display antibodies using libraries derived from chicken immunoglobulin genes" J Immunol Methods 1995 Oct 12;186( 1 ):125-35; Jones C RT al. "Current trends in molecular recognition and bioseparation" J
Chromatogr A 1995 Jul 14;707( 1 ):3-22; Deng SJ et al. "Basis for selection of improved carbohydrate-binding single-chain antibodies from synthetic gene libraries"
Proc Natl Acad Sci U S A 1995 May 23;92(11):4992-6; and Deng SJ et al. "Selection of antibody single-chain variable fragments with improved carbohydrate binding by phage display" J Biol Chem 1994 Apr 1;269(13):9533-8, which are incorporated herein by reference.
The following sections relate to Candidate Marker Examples (first section) and to Experimental Data for these Marker Examples (second section). It should be noted that Table numbering is restarted within each section.
CANDIDATE MARKER EXAMPLES SECTION
This Section relates to Examples of sequences according to the present invention, including illustrative methods of selection thereof.
Description of the methodology undertaken to uncover the biomolecular sequences of the present invention Human ESTs and cDNAs were obtained from GenBank versions 136 (June 15, 2003 ftp.ncbi.nih.gov/genbank/release.notes/gb136.release.notes); NCBI genome assembly of April 2003; Refseq sequences from June 2003; Genbank version 139 (December 2003);
Human Genome from NCBI (Build 34) (from Oct 2003); and Ref~eq sequences from December 2003;
and from the LifeSeq library of Incyte Corporation (ES Ts only; Wilmington, DE, USA). With regard to GenBank sequences, the human EST sequences from the EST (GBEST) section and the human mRNA sequences from the primate (GBPRI) section were used; also the human nucleotide Refseq mRNA sequences were used (see for example www.ncbi.nlm.nih.gov/Genbank/GenbankOverview.html and for a reference to the EST section, see www.ncbi.nlm.nih.gov/dbEST/; a general reference to dbEST, the EST
database in GenBank, may be found in Boguski et al, Nat Genet. 1993 Aug;4(4):332-3; all of which are hereby incorporated by reference as if fully set forth herein).
Novel splice variants were predicted using the LEADS clustering and assembly system as described in Sorek, R., Ast, G. & Graur, D. Alu-containing exons are alternatively spliced.
Genome Res 12, 1060-7 (2002); US patent No: 6,625,545; and U.S. Pat. Appl. No.
10/426,002, published as US20040101876 on May 27 2004; all of which are hereby incorporated by reference as if fully set forth herein. Briefly, the software cleans the expressed sequences from repeats, vectors and immunoglobulins. It then aligns the expressed sequences to the genome taking alternatively splicing into account and clusters overlapping expressed sequences into "clusters" that represent genes or partial genes.
These were annotated using the GeneCarta (Compugen, Tel-Aviv, Israel) platform. The GeneCarta platform includes a rich pool of annotations, sequence information (particularly of spliced sequences), chromosomal information, alignments, and additional information such as SNPs, gene ontology terms, expression profiles, functional analyses, detailed domain structures, known and predicted proteins and detailed homology reports.
A brief explanation is provided with regard to the method of selecting the candidates.
However, it should noted that this explanation is provided for descriptive purposes only, and is not intended to be limiting in any way. The potential markers were identified by a computational process that was designed to find genes and/or their splice variants that are over-expressed in tumor tissues, by using databases of expressed sequences. Various parameters related to the information in the EST libraries, determined according to a manual classification process, were used to assist in locating genes and/or splice variants thereof that are over-expressed in cancerous tissues. The detailed description of the selection method is presented in Example 1 below. The cancer biomarkers selection engine and the following wet validation stages are schematically summarized in Figure 1.
Identification of differentially expressed gene products - Algorithm In order to distinguish between differentially expressed gene products and constitutively expressed genes (i.e., house keeping genes ) an algorithm based on an analysis of frequencies was configured. A specific algorithm for identification of transcripts over expressed in cancer is described hereinbelow.
Dry analysis Library annotation - EST libraries are manually classified according to:
(i) Tissue origin (ii) Biological source - Examples of frequently used biological sources for construction of EST libraries include cancer cell-lines; normal tissues; cancer tissues; fetal tissues; and others such as normal cell lines and pools of normal cell-lines, cancer cell-lines and combinations thereof. . A specific description of abbreviations used below with regard to these tissues/cell lines etc is given above.
(iii) Protocol of library construction - various methods are known in the art for library construction including normalized library construction;
non-normalized library construction; subtracted libraries; ORESTES and others. It will be appreciated that at times the protocol of library construction is not indicated.
The following rules were followed:
EST libraries originating from identical biological samples are considered as a single library.
EST libraries which included above-average levels of contamination, such as DNA
contamination for example, were eliminated. The presence of such contamination was determined as follows. For each library, the number of unspliced ESTs that are not fully contained within other spliced sequences was counted. If the percentage of such sequences (as compared to all other sequences) was at least 4 standard deviations above the average for all libraries being analyzed, this library was tagged as being contaminated and was eliminated from further consideration in the below analysis (see also Sorek, R. & Safer, H.M. A novel algorithm for computational identification of contaminated EST libraries. Nucleic Acids Res 31, 1067-74 (2003)for further details).
Clusters (genes) having at least five sequences including at least two sequences from the tissue of interest were analyzed. Splice variants were identified by using the LEADS software package as described above.
Identification of genes over expressed in cancer.
Two different scoring algorithms were developed.
Libraries score -candidate sequences which are supported by a number of cancer libraries, are more likely to serve as specific and effective diagnostic markers.
The basic algorithm - for each cluster the number of cancer and normal libraries contributing sequences to the cluster was counted. Fisher exact test was used to check if cancer libraries are significantly over-represented in the cluster as compared to the total number of cancer and normal libraries.
Library counting: Small libraries (e.g., less than 1000 sequences) were excluded from consideration unless they participate in the cluster. For this reason, the total number of libraries is actually adjusted for each cluster.
Clones no. score - Generally, when the number of ESTs is much higher in the cancer libraries relative to the normal libraries it might indicate actual over-expression.
The algorithm -Clone counting : For counting EST clones each library protocol class was given a weight based on our belief of how much the protocol reflects actual expression levels:
(i) non-normalized : 1 (ii) normalized : 0.2 (iii) all other classes : 0.1 Clones number score - The total weighted number of EST clones from cancer libraries was compared to the EST clones from normal libraries. To avoid cases where one library contributes to the majority of the score, the contribution of the library that gives most clones for a given cluster was limited to 2 clones.
The score was computed as C+1 C
n+1 N
where:
c - weighted number of "cancer" clones in the cluster.
C- weighted number of clones in all "cancer" libraries.
n - weighted number of "normal" clones in the cluster.
N- weighted number of clones in all "normal" libraries.
Clones number score significance - Fisher exact test was used to check if EST
clones from cancer libraries are significantly over-represented in the cluster as compared to the total number of EST clones from cancer and normal libraries.
Two search approaches were used to find either general cancer-specific candidates or tumor specific candidates.
~ Libraries/sequences originating from tumor tissues are counted as well as libraries originating from cancer cell-lines ("normal" cell-lines were ignored).
~ Only libraries/sequences originating from tumor tissues are counted Identification of tissue specific genes For detection of tissue specific clusters, tissue libraries/sequences mere compared to the total number of libraries/sequences in cluster. Similar statistical tools to those described in above were employed to identify tissue specific genes. Tissue abbreviations are the same as for cancerous tissues, but are indicated with the header "normal tissue".
The algorithm - for each tested tissue T and for each tested cluster the following were examined:
1. Each cluster includes at least 2 libraries from the tissue T. At least 3 clones (weighed - as described above) from tissue T in the cluster; and 2. Clones from the tissue T are at least 40 % from all the clones participating in the tested cluster Fisher exact test Rvalues were computed both for library and weighted clone counts to check that the counts are statistically significant.
Identification of splice variants over expressed in cancer of clusters which are not over expressed in cancer Cancer-specific splice variants containing a unique region were identified.
Identification of unique sequence regions in splice variants A Region is defined as a group of adjacent exons that always appear or do not appear together in each splice variant.
A "segment" (sometimes referred also as "seg" or "node") is defined as the shortest contiguous transcribed region without known splicing inside.
, Only reliable ESTs were considered for region and segment analysis. An EST
was defined as unreliable i~
(i) Unspliced;
(ii) Not covered by RNA;
(iii) Not covered by spliced ESTs; and (iv) Alignment to the genome ends in proximity of long poly-A stretch or starts in proximity of long poly-T stretch.
Only reliable regions were selected for further scoring. Unique sequence regions were considered reliable i~
(i) Aligned to the genome; and (ii) Regions supported by more than 2 ESTs.
The algorithm Each unique sequence region divides the set of transcripts into 2 groups:
(i) Transcripts containing this region (group TA).
(ii) Transcripts not containing this region (group TB).
The set of EST clones of every cluster is divided into 3 groups:
(i) Supporting (originating from) transcripts of group TA (S 1 ).
(ii) Supporting transcripts of group TB (S2).
(iii) Supporting transcripts from both groups (S3).
Library and clones number scores described above were given to S1 group.
Fisher Exact Test P-values were used to check i~
S 1 is significantly enriched by cancer EST clones compared to S2; and S1 is significantly enriched by cancer EST clones compared to cluster background (S1+S2+S3).
Identification of unique sequence regions and division of the group of transcripts accordingly is illustrated in Figure 2. Each of these unique sequence regions corresponds to a segment, also termed herein a "node".
Region 1: conunon to all transcripts, thus it is preferably not considered for determining differential expression between variants; Region 2: specific to Transcript 1;
Region 3: specific to Transcripts 2+3; Region 4: specific to Transcript 3; Region 5: specific to Transcripts 1 and 2;
Region 6: specific to Transcript 1.
Identification of cancer specific splice variants of genes over expressed in cancer A search for EST supported (no mRNA) regions for genes o~
(i) known cancer markers (ii) Genes shown to be over-expressed in cancer in published micro-array experiments.
Reliable EST supported-regions were defined as supported by minimum of one of the following:
(i) 3 spliced ESTs; or (ii) 2 spliced ESTs from 2 libraries;
(iii) 10 unspliced ESTs from 2 libraries, or (iv) 3libraries.
Actual Marker Examples The following examples relate to specific actual marker examples. It should be noted that Table numbering is restarted within each example related to a particular Cluster, as indicated by the titles below.
EXPERIMENTAL EXAMPLES SECTION
This Section relates to Examples describing experiments involving these sequences, and illustrative, norrlimiting examples of methods, assays and uses thereof. The materials and experimental procedures are explained first, as all experiments used them as a basis for the work I O that was performed.
The markers of the present invention were tested with regard to their expression in various cancerous and norrcancerous tissue samples. A description of the samples used in the panel is provided in Table 1 below. A description of the samples used in the normal tissue panel is provided in Table 2 below. Tests were then performed as described in the "Materials and Experimental Procedures" section below.
Table 1: Tissue samples in testing panel Lot Sample numbe Gra name r ource Tissue Pathology de gender/age 2-A-Pap ILS- Papillary Adeno G2 1408 BS ovary adenocarcinoma 2 53/F
3-A-Pap ILS- Papillary Adeno G2 1431 BS ovary adenocarcinoma 2 52/F
4-A-Pap CystAdeno ILS- Papillary G2 7286 BS ovary cystadenocarcinoma2 50/F
1-A-Pap ILS- BS ovary Papillary 3 73/F
Adeno G3 1406 adenocarcinoma 14-B-AdenoA50111BioChai G2 1 n ovary Adenocarcinoma 2 41/F
5-G-Adeno 99-12- Adenocarcinoma G3 6432 GOG ovary (Stage3C) 3 46/F
6-A-Adeno A0106 BS ovary adenocarcinoma 3 51/F
IND-7-A-Adeno 00375 BS ovary adenocarcinoma 3 59/F
A50111BioChai 8-B-Adeno 3 n ovary adenocarcinoma 3 60/F
9-G-Adeno 99-06- Adenocarcinoma G3 6901 GOG ovary (maybe serous) 3 84/F
10-B-AdenoA40706Biochai G3 9 n ovary Adenocarcinoma 3 60/F
11-B-AdenoA40706Biochai G3 8 n ovary Adenocarcinoma 3 49/F
12-B-AdenoA40602Biochai G3 3 n ovary Adenocarcinoma 3 45/F
13-G-Adeno94-05- right Metastasis G3 7603 GOG ovary adenocarcinoma 3 67/F
15-B-AdenoA40706BioChai G3 5 n ovary Carcinoma 3 27/F
10903 Clontec 16-Ct-Adeno7 h ovary Carcinoma NOS F
Mucinous 22-A-Muc cystadenocarcinoma CystAde A0139 BS ovary (Stage1 C) 2 72/F
Mucinous 21-G- Muc 95-10- cystadenocarcinoma CystAde 6020 GOG ovary (Stage2) 2-3 44/F
Mucinous 23-A-Muc VNM- cystadenocarcinoma CystAde 00187 BS ovary with low malignant3 45/F
17-B-Muc A50408BioChai Mucinous Adeno G3 4 n ovary adenocarcinoma 3 51/F
18-B-Muc A50408BioChai Mucinous Adeno G3 3 n ovary adenocarcinoma 3 45/F
19- B-Muc A50408BioChai Mucinous Adeno G3 5 n ovary adenocarcinoma 34/F
20- A-Pap USA- Papillary mucinous Muc CystAde00273 BS ovary cystadenocarcinoma 45/F
33-B-Pa p Sero CystAdeA50317BioChai Serous papillary G1 5 n ovary cystadenocarcinoma1 41/F
25-A-Pap Papillary serous Sero Adeno adenocarcinoma G3 N0021 BS ovary (StageT3CN1 3 55/F
MX) 24-G-Pap 2001-Sero Adeno07- Papillary serous G3 6801 GOG ovary adenocarcinoma 3 68/F
30-G-Pap 2001-Sero Adeno08- Papillary serous G3 6011 GOG ovary carcinoma (Stage1C)3 72/F
70-G-Pap Sero Adeno95-08- Papillary serous G3 6069 GOG ovary adenocarcinoma 3 F
31-B-Pa p Sero CystAdeA50317BioChai Serous papillary G3 ~ 6 n ovary cystadenocarcinoma3 52/F
32-G-Pap 93-09-GOG ovary Serous papillary3 F
Sero CystAde4901 cystadenocarcinoma Papillary serous 66-G-Pap 2000- carcinoma (metastais Sero Adeno01- of primary G3 SIV 6413 GOG ovary peritoneum) F
(Stage4) 2001- Serous 29-G-Sero 12- right adenocarcinoma Adeno G3 6035 GOG ovary (Stage3A) 3 50/F
Mixed epithelial cystadenocarcinoma with mucinous, endometrioid, 41-G-Mix squamous and Sero/Muc/End98-03- papillary serous o G2 6803 GOG ovary (Stage2) 2 38 Papillary serous and endometrioid 40-G-Mix 95-11- ovary,endcystadenocarcinoma Sero/Endo 6006 GOG ometrium(Stage3C) 2 49/F
2002- Mixed serous and 37-G-Mix 05- endometrioid Sero/Endo 6513 GOG ovary adenocarcinoma 3 56/F
Mixed serous and 2002- endometrioid 38-G-Mix 05- adenocarcinoma of Sero/Endo 6509 GOG ovary mullerian (Stage3C)3 64/F
2001- Mixed serous and 39--G-Mix 12- endometrioid Sero/Endo 6037 GOG ovary adenocarcinoma 3 F
36-G-Endo 09- Endometrial Adeno G1-26621 GOG ovary adenocarcinoma 1-2 69/F
35-G-Endo 94-08- right Endometrioid Adeno G2 7604 GOG ovary adenocarcinoma 2 39/F
Papillary 34-G-Pap endometrioid Endo Adeno95-04- adenocarcinoma G3 2002 GOG ovary (Stage3C) 3 68/F
43-G-Clear10- Clear cell cell Adeno6002 GOG ovary adenocarcinoma 3 74/F
2001- Clear cell 44-G-Clear07- adenocarcinoma cell Adeno6084 GOG ovary (Stage3A) 73/F
Epithelial adenocarcinoma of 42-G-Adeno98-08- borderline borderline6001 GOG ovary malignancy 46/F
59-G-Sero CysAdenoFibr98-12- Serous oma 6401 GOG ovary CysAdenoFibroma 77/F
Serous 63-G-Sero 2000- CysAdenoFibroma of CysAdenoFibr10- borderline oma 6620 GOG ovary malignancy 71/F
64-G-Ben Sero 99-06- Bengin Serous CysAdenoma6039 GOG ovary CysAdenoma 57/F
56-G-Ben 99-01-GOG left Bengin mucinus 46/F
ovary Muc 6407 cysadenoma CysAdeno a n Muc 99-10- Bengin mucinus CysAdenoma6442 GOG ovary cysadenoma 32/F
60-G- Muc 99-01- Mucinous CysAdenoma6043 GOG ovary Cysadenoma 40/F
61-G- Muc 99-07- Mucinous CysAdenoma6011 GOG ovary Cysadenoma 63/F
Endometriom97-11- right a 6320 GOG ovary Endometrioma 41/F
57-B- A40706BioChai Thecoma 6 n ovary Thecoma 56/F
Struma 58-CG-Stru ovary/monodermal teratoma CG-1771chilovovary teratoma 58/F
A50111BioChai Normal (matched 50-B-N 4 n ovary tumor A501113) 60/F
A50111BioChai Normal (matched 49-B-N 2 n ovary tumor A501111 41 /F
M14 ) 07- Normal (matched 69-G-N 6801 GOG ovary tumor 2001-07-6801 68/F
M24 N ) 05- Normal (matched 67-G-N 509N GOG ovary tumor 2002-05-6509) 64/F
98-03- Normal (matched 51-G-N G803N GOG ovary tumor 98-03-6803) 38/F
52-G-N 98-08-GOG ovary Normal (matched 46/F
6001 tumor 98-08-6001 N ) 99-01- Normal (matched 68-G-N G407N GOG ovary bengin 99-01-6407) 46/F
01- Normal (matched 72-G-N G413N GOG ovary tumor 2000-01-6413) F
98-12- Normal (matched 73-G-N 6401 GOG ovary tumor 98-12-6401 77/F
M59 N ) 97-11- Normal (matched 74-G-N G320N GOG ovary tumor 97-116320) 41/F
99-01- Normal (matched 75-G-N G043N GOG ovary tumor 99-01-6043) 40/F
A50327BioChai 45-B-N 4 n ovary Normal PM 41/F
A50408BioChai 46-B-N 6 n ovary Normal PM 41/F
A50408BioChai 48-B-N 7 n ovary Normal PM 51/F
061 Normal (CLOSED
47-Am-N A mbion ovary HEAD) 16/F
CG-71-CG-N 188-7 Ichilovovary Normal PM 49/F
Table 2: Tissue samples in normal panel:
Lot ource issue Pathology Sex/Age no.
1-Am-Colon (C71 071PIOBmbion Colon PM F/43 ) 2-B-Colon (C69) A411078BiochainColon PM-Pool M&F
of 10 3-CI-Colon (C70)I I ClontechColon PM-Pool M&F
10101 of 3 4-Am-Small Intestine091P0201mbion Small IntestinePM M/75 5-B-Small IntestineA501 BiochainSmall IntestinePM M/63 6-B-Rectum A605138BiochainRectum PM M/25 7-B-Rectum A610297BiochainRectum PM M/24 8-B-Rectum A610298BiochainRectum PM M/27 9-Am-Stomach 110P04Ambion Stomach PM M/16 10-B-Stomach A501159BiochainStomach PM M/24 11-B-Esophagus A603814BiochainEsophagus PM M/26 12-B-Esophagus A603813BiochainEsophagus PM M/41 13-Am-Pancreas 071P25Cmbion Pancreas PM M/25 14-CG-Pancreas CG-255-2IchilovPancreas PM M/75 15-B-Lung A409363BiochainLung PM F/26 16-Am-Lung (L93)111P0103mbion Lung PM F/61 17-B-Lung (L92) A503204BiochainLung PM M/28 18-Am-Ovary (047)061P43Ambion Ovary PM F/16 19-B-Ovary (048)A504087BiochainOvary PM F/51 20-B-Ovary (046)A504086BiochainOvary PM F/41 21-Am-Cervix lOIPOlOIAmbion Cervix PM F/40 22-B-Cervix A408211BiochainCervix PM F/36 23-B-Cervix A504089BiochainCervix PM-Pool M8~F
of 5 24-B-Uterus A411074BiochainUterus PM-Pool M&F
of 10 25-B-Uterus A409248BiochainUterus PM F/43 26-B-Uterus A504090BiochainUterus PM-Pool M&F
of 5 27-B-Bladder A501157BiochainBladder PM M/29 28-Am-Bladder 071P02Cmbion Bladder PM M/20 29-B-Bladder A504088BiochainBladder PM-Pool M8~F
of 5 30-Am-Placenta 021P33Ambion Placenta PB F/33 31-B-Placenta A410165BiochainPlacenta PB F/26 32-B-Placenta A411073BiochainPlacenta PB-Pool M&F
of 5 33-B-Breast (B59)A607155BiochainBreast PM F/36 34-Am-Breast 26486 mbion Breast PM F/43 (B63) 35-Am-Breast 23036 mbion Breast PM F/57 (B64) 36-CI-Prostate 1070317 ClontechProstate PB-Pool M&F
(P53) of 47 37-Am-Prostate 061 P04Ambion Prostate PM M/47 (P42) 38-Am-Prostate 25955 mbion Prostate PM M/62 (P59) 39-Am-Testis 111P0104Ambion estis PM M/25 40-B-Testis A411147 Biochainestis PM M/74 41-CI-Testis 1110320 Clontechestis PB-Pool M&F
of 45 42-CG-Adrenal CG-184-101chilovdrenal PM F/81 43-B-Adrenal A610374 Biochaindrenal PM F/83 44-B-Heart A411077 BiochainHeart PB-Pool M&F
of 5 45-CG-Heart CG-255-9IchilovHeart PM M/75 46-CG-Heart CG-227-1IchilovHeart PM F/36 47-Am-Liver 081P0101mbion Liver PM M/64 48-CG-Liver CG-93-3 IchilovLiver PM F/19 49-CG-Liver CG-124-4IchilovLiver PM F/34 50-CI-BM 1110932 ClontechBone MarrowPM-Pool M&F
of 8 51-CGEN-Blood WBC#5 CGEN Blood M
15-B-AdenoA40706BioChai G3 5 n ovary Carcinoma 3 27/F
10903 Clontec 16-Ct-Adeno7 h ovary Carcinoma NOS F
Mucinous 22-A-Muc cystadenocarcinoma CystAde A0139 BS ovary (Stage1 C) 2 72/F
Mucinous 21-G- Muc 95-10- cystadenocarcinoma CystAde 6020 GOG ovary (Stage2) 2-3 44/F
Mucinous 23-A-Muc VNM- cystadenocarcinoma CystAde 00187 BS ovary with low malignant3 45/F
17-B-Muc A50408BioChai Mucinous Adeno G3 4 n ovary adenocarcinoma 3 51/F
18-B-Muc A50408BioChai Mucinous Adeno G3 3 n ovary adenocarcinoma 3 45/F
19- B-Muc A50408BioChai Mucinous Adeno G3 5 n ovary adenocarcinoma 34/F
20- A-Pap USA- Papillary mucinous Muc CystAde00273 BS ovary cystadenocarcinoma 45/F
33-B-Pa p Sero CystAdeA50317BioChai Serous papillary G1 5 n ovary cystadenocarcinoma1 41/F
25-A-Pap Papillary serous Sero Adeno adenocarcinoma G3 N0021 BS ovary (StageT3CN1 3 55/F
MX) 24-G-Pap 2001-Sero Adeno07- Papillary serous G3 6801 GOG ovary adenocarcinoma 3 68/F
30-G-Pap 2001-Sero Adeno08- Papillary serous G3 6011 GOG ovary carcinoma (Stage1C)3 72/F
70-G-Pap Sero Adeno95-08- Papillary serous G3 6069 GOG ovary adenocarcinoma 3 F
31-B-Pa p Sero CystAdeA50317BioChai Serous papillary G3 ~ 6 n ovary cystadenocarcinoma3 52/F
32-G-Pap 93-09-GOG ovary Serous papillary3 F
Sero CystAde4901 cystadenocarcinoma Papillary serous 66-G-Pap 2000- carcinoma (metastais Sero Adeno01- of primary G3 SIV 6413 GOG ovary peritoneum) F
(Stage4) 2001- Serous 29-G-Sero 12- right adenocarcinoma Adeno G3 6035 GOG ovary (Stage3A) 3 50/F
Mixed epithelial cystadenocarcinoma with mucinous, endometrioid, 41-G-Mix squamous and Sero/Muc/End98-03- papillary serous o G2 6803 GOG ovary (Stage2) 2 38 Papillary serous and endometrioid 40-G-Mix 95-11- ovary,endcystadenocarcinoma Sero/Endo 6006 GOG ometrium(Stage3C) 2 49/F
2002- Mixed serous and 37-G-Mix 05- endometrioid Sero/Endo 6513 GOG ovary adenocarcinoma 3 56/F
Mixed serous and 2002- endometrioid 38-G-Mix 05- adenocarcinoma of Sero/Endo 6509 GOG ovary mullerian (Stage3C)3 64/F
2001- Mixed serous and 39--G-Mix 12- endometrioid Sero/Endo 6037 GOG ovary adenocarcinoma 3 F
36-G-Endo 09- Endometrial Adeno G1-26621 GOG ovary adenocarcinoma 1-2 69/F
35-G-Endo 94-08- right Endometrioid Adeno G2 7604 GOG ovary adenocarcinoma 2 39/F
Papillary 34-G-Pap endometrioid Endo Adeno95-04- adenocarcinoma G3 2002 GOG ovary (Stage3C) 3 68/F
43-G-Clear10- Clear cell cell Adeno6002 GOG ovary adenocarcinoma 3 74/F
2001- Clear cell 44-G-Clear07- adenocarcinoma cell Adeno6084 GOG ovary (Stage3A) 73/F
Epithelial adenocarcinoma of 42-G-Adeno98-08- borderline borderline6001 GOG ovary malignancy 46/F
59-G-Sero CysAdenoFibr98-12- Serous oma 6401 GOG ovary CysAdenoFibroma 77/F
Serous 63-G-Sero 2000- CysAdenoFibroma of CysAdenoFibr10- borderline oma 6620 GOG ovary malignancy 71/F
64-G-Ben Sero 99-06- Bengin Serous CysAdenoma6039 GOG ovary CysAdenoma 57/F
56-G-Ben 99-01-GOG left Bengin mucinus 46/F
ovary Muc 6407 cysadenoma CysAdeno a n Muc 99-10- Bengin mucinus CysAdenoma6442 GOG ovary cysadenoma 32/F
60-G- Muc 99-01- Mucinous CysAdenoma6043 GOG ovary Cysadenoma 40/F
61-G- Muc 99-07- Mucinous CysAdenoma6011 GOG ovary Cysadenoma 63/F
Endometriom97-11- right a 6320 GOG ovary Endometrioma 41/F
57-B- A40706BioChai Thecoma 6 n ovary Thecoma 56/F
Struma 58-CG-Stru ovary/monodermal teratoma CG-1771chilovovary teratoma 58/F
A50111BioChai Normal (matched 50-B-N 4 n ovary tumor A501113) 60/F
A50111BioChai Normal (matched 49-B-N 2 n ovary tumor A501111 41 /F
M14 ) 07- Normal (matched 69-G-N 6801 GOG ovary tumor 2001-07-6801 68/F
M24 N ) 05- Normal (matched 67-G-N 509N GOG ovary tumor 2002-05-6509) 64/F
98-03- Normal (matched 51-G-N G803N GOG ovary tumor 98-03-6803) 38/F
52-G-N 98-08-GOG ovary Normal (matched 46/F
6001 tumor 98-08-6001 N ) 99-01- Normal (matched 68-G-N G407N GOG ovary bengin 99-01-6407) 46/F
01- Normal (matched 72-G-N G413N GOG ovary tumor 2000-01-6413) F
98-12- Normal (matched 73-G-N 6401 GOG ovary tumor 98-12-6401 77/F
M59 N ) 97-11- Normal (matched 74-G-N G320N GOG ovary tumor 97-116320) 41/F
99-01- Normal (matched 75-G-N G043N GOG ovary tumor 99-01-6043) 40/F
A50327BioChai 45-B-N 4 n ovary Normal PM 41/F
A50408BioChai 46-B-N 6 n ovary Normal PM 41/F
A50408BioChai 48-B-N 7 n ovary Normal PM 51/F
061 Normal (CLOSED
47-Am-N A mbion ovary HEAD) 16/F
CG-71-CG-N 188-7 Ichilovovary Normal PM 49/F
Table 2: Tissue samples in normal panel:
Lot ource issue Pathology Sex/Age no.
1-Am-Colon (C71 071PIOBmbion Colon PM F/43 ) 2-B-Colon (C69) A411078BiochainColon PM-Pool M&F
of 10 3-CI-Colon (C70)I I ClontechColon PM-Pool M&F
10101 of 3 4-Am-Small Intestine091P0201mbion Small IntestinePM M/75 5-B-Small IntestineA501 BiochainSmall IntestinePM M/63 6-B-Rectum A605138BiochainRectum PM M/25 7-B-Rectum A610297BiochainRectum PM M/24 8-B-Rectum A610298BiochainRectum PM M/27 9-Am-Stomach 110P04Ambion Stomach PM M/16 10-B-Stomach A501159BiochainStomach PM M/24 11-B-Esophagus A603814BiochainEsophagus PM M/26 12-B-Esophagus A603813BiochainEsophagus PM M/41 13-Am-Pancreas 071P25Cmbion Pancreas PM M/25 14-CG-Pancreas CG-255-2IchilovPancreas PM M/75 15-B-Lung A409363BiochainLung PM F/26 16-Am-Lung (L93)111P0103mbion Lung PM F/61 17-B-Lung (L92) A503204BiochainLung PM M/28 18-Am-Ovary (047)061P43Ambion Ovary PM F/16 19-B-Ovary (048)A504087BiochainOvary PM F/51 20-B-Ovary (046)A504086BiochainOvary PM F/41 21-Am-Cervix lOIPOlOIAmbion Cervix PM F/40 22-B-Cervix A408211BiochainCervix PM F/36 23-B-Cervix A504089BiochainCervix PM-Pool M8~F
of 5 24-B-Uterus A411074BiochainUterus PM-Pool M&F
of 10 25-B-Uterus A409248BiochainUterus PM F/43 26-B-Uterus A504090BiochainUterus PM-Pool M&F
of 5 27-B-Bladder A501157BiochainBladder PM M/29 28-Am-Bladder 071P02Cmbion Bladder PM M/20 29-B-Bladder A504088BiochainBladder PM-Pool M8~F
of 5 30-Am-Placenta 021P33Ambion Placenta PB F/33 31-B-Placenta A410165BiochainPlacenta PB F/26 32-B-Placenta A411073BiochainPlacenta PB-Pool M&F
of 5 33-B-Breast (B59)A607155BiochainBreast PM F/36 34-Am-Breast 26486 mbion Breast PM F/43 (B63) 35-Am-Breast 23036 mbion Breast PM F/57 (B64) 36-CI-Prostate 1070317 ClontechProstate PB-Pool M&F
(P53) of 47 37-Am-Prostate 061 P04Ambion Prostate PM M/47 (P42) 38-Am-Prostate 25955 mbion Prostate PM M/62 (P59) 39-Am-Testis 111P0104Ambion estis PM M/25 40-B-Testis A411147 Biochainestis PM M/74 41-CI-Testis 1110320 Clontechestis PB-Pool M&F
of 45 42-CG-Adrenal CG-184-101chilovdrenal PM F/81 43-B-Adrenal A610374 Biochaindrenal PM F/83 44-B-Heart A411077 BiochainHeart PB-Pool M&F
of 5 45-CG-Heart CG-255-9IchilovHeart PM M/75 46-CG-Heart CG-227-1IchilovHeart PM F/36 47-Am-Liver 081P0101mbion Liver PM M/64 48-CG-Liver CG-93-3 IchilovLiver PM F/19 49-CG-Liver CG-124-4IchilovLiver PM F/34 50-CI-BM 1110932 ClontechBone MarrowPM-Pool M&F
of 8 51-CGEN-Blood WBC#5 CGEN Blood M
52-CGEN-Blood WBC#4 CGEN Blood M
53-CGEN-Blood WBC#3 CGEN Blood M
54-CG-Spleen CG-267 IchilovSpleen PM F/25 55-CG-Spleen 111P0106Bmbion Spleen PM M/25 56-CG-Spleen A409246 BiochainSpleen PM F/12 56-CG-Thymus CG-98-7 Ichilovhymns PM F/28 58-Am-Thymus lOIPOIOIAmbion hymns PM M/14 59-B-Thymus A409278 Biochainhymns PM M/28 60-B-Thyroid A610287 Biochainhyroid PM M/27 61-B-Thyroid A610286 Biochainhyroid PM M/24 62-CG-Thyroid CG-119-2Ichilovhyroid PM F/66 63-CI-Salivary 1070319 ClontechSalivary PM-Pool M&F
Gland Gland of 24 64-Am-Kidney 111PO1O1Bmbion Kidney PM-Pool M8~F
of 14 65-CI-Kidney 1110970 ClontechKidney PM-Pool M&F
of 14 66-B-Kidney A411080 BiochainKidney PM-Pool M&F
of 5 67-CG-CerebellumCG-183-5IchilovCerebellum PM M/74 68-CG-CerebellumCG-212-5IchilovCerebellum PM M/54 69-B-Brain A411322 BiochainBrain PM M/28 70-CI-Brain 1120022 ClontechBrain PM-Pool M&F
of 2 71-B-Brain A411079 BiochainBrain PM-Pool M&F
of 2 72-CG-Brain CG-151-IIchilovBrain PM F/86 73-Am-Skeletal 1O1P013Ambion Skeletal PM F/28 Muscle Muscle 74-CI-Skeletal 1061038 ClontechSkeletal PM-Pool M&F
Muscle Muscle of 2 Materials and Experimental Procedures RNA preparation - RNA was obtained from Clontech (Franklin Lakes, NJ USA
07417, www.clontech.com), BioChain Inst. Inc. (Hayward, CA 94545 USA
www.biochain.com), ABS
(Wilmington, DE 19801, USA, http://www.absbioreagents.com) or Ambion (Austin;
USA, http://www.ambion.com). Alternatively, RNA was generated from tissue samples using TRI-Reagent (Molecular Research Center), according to Manufacturer's instructions. Tissue and RNA samples were obtained from patients or from postmortem. Total RNA samples were treated with DNaseI (Ambion) and purified using RNeasy columns (Qiagen).
RT PCR - Purified RNA (1 pg) was mixed with I50 ng Random Hexamer primers (Invitrogen) and 500 pM dNTP in a total volume of 15.6 pl. The mixture was incubated for 5 min at 65 °C and then quickly chilled on ice. Thereafter, 5 p.l of SX
SuperscriptII first strand buffer (Invitrogen), 2.4p,1 O.1M DTT and 40 units RNasin (Promega) were added, and the mixture was incubated for 10 min at 25 °C, followed by further incubation at 42 °C for 2 min.
Then, 1 p l (200units) of SuperscriptII (Invitrogen) was added and the reaction (final volume of 25p1) was incubated for 50 min at 42 °C and then inactivated at 70 °C for ISmin. The resulting cDNA was diluted 1:20 in TE buffer (10 mM Tris pH=8, 1 mM EDTA pH=8).
Real-Time RT-PCR analysis- cDNA (Spl), prepared as described above, was used as a template in Real-Time PCR reactions using the SYBR Green I assay (PE Applied Biosystem) with specific primers and UNG Enzyme (Eurogentech or ABI or Roche). The amplification was effected as follows: 50 "C for 2 min, 95 °C for 10 min, and then 40 cycles of 95 "C for l5sec, followed by 60 °C for 1 min. Detection was performed by using the PE
Applied Biosystem SDS
7000. The cycle in which the reactions achieved a threshold level (Ct) of fluorescence was registered and was used to calculate the relative transcript quantity in the RT reactions. The relative quantity was calculated using the equation Q=efficiency~-~'. The efficiency of the PCR
reaction was calculated from a standard curve, created by using serial dilutions of several reverse transcription (RT) reactions. To minimize inherent differences in the RT reaction, the resulting relative quantities were normalized to the geometric mean of the relative quantities of several housekeeping (HSKP) genes. Schematic summary of quantitative real-time PCR
analysis is presented in Figure 3. As shown, the x axis shows the cycle number. The CT =
Threshold Cycle point, which is the cycle that the amplification curve crosses the fluorescence threshold that was set in the experiment. This point is a calculated cycle number in which PCR
products signal is above the background level (passive dye ROX) and still in the Geometric/Exponential phase (as shown, once the level of fluorescence crosses the measurement threshold, it has a geometrically increasing phase, during which measurements are most accurate, followed by a linear phase and a plateau phase; for quantitative measurements, the latter two phases do not provide accurate measurements). The taxis shows the normalized reporter fluorescence. It should be noted that this type of analysis provides relative quantification.
The sequences of the housekeeping genes measured in all the examples on ovarian cancerpanel were as follows:
SDHA (GenBank Accession No.1VM-004168) SDHA Forward primer: TGGGAACAAGAGGGCATCTG
SDHA Reverse primer: CCACCACTGCATCAAATTCATG
SDHA-amplicon TGGGAACAAGAGGGCATCTGCTAAAGTTTCAGATTCCATTTCTGCTCAGTATCCAGT
AGTGGATCATGAATTTGATGCAGTGGTGG
PBGD (GenBank Accession No. BC019323), PBGD Forward primer: TGAGAGTGATTCGCGTGGG
PBGD Reverse primer: CCAGGGTACGAGGCTTTCAAT
PBGD-amplicon:
TGAGAGTGATTCGCGTGGGTACCCGCAAGAGCCAGCTTGCTCGCATACAGACGGAC
AGTGTGGTGGCAACATTGAAAGCCTCGTACCCTGG
HPRT1 (GenBank Accession No. NM 000194), HPRT1 Forward primer: TGACACTGGCAAAACAATGCA
HPRT1 Reverse primer: GGTCCTTTTCACCAGCAAGCT .
HPRT1-amplicon:
TGACACTGGCAAAACAATGCAGACTTTGCTTTCCTTGGTCAGGCAGTATAATCCAA
AGATGGTCAAGGTCGCAAGCTTGCTGGTGAAAAGGACC
GAPDH (GenBank Accession No. BC026907) GAPDH Forward primer: TGCACCACCAACTGCTTAGC
GAPDH Reverse primer: CCATCACGCCACAGTTTCC
GAPDH-amplicon TGCACCACCAACTGCTTAGCACCCCTGGCCAAGGTCATCCATGACAACTTTGGTATC
GTGGAAGGACTCATGACCACAGTCCATGCCATCACTGCCACCCAGAAGACTGTGGA
TGG
The sequences of the housekeeping genes measured in all the examples on normal tissue samples panel were as follows:
RPL19 (GenBank Accession No. NM_000981), RPL19 Forward primer: TGGCAAGAAGAAGGTCTGGTTAG
RPL19 Reverse primer: TGATCAGCCCATCTTTGATGAG
RPL19 -amplicon:
TGGCAAGAAGAAGGTCTGGTTAGACCCCAATGAGACCAATGAAATCGCCAATGCCA
ACTCCCGTCAGCAGATCCGGAAGCTCATCAAAGATGGGCTGATCA
TATA box (GenBank Accession No. NM 003194), TATA box Forward primer : CGGTTTGCTGCGGTAATCAT
TATA box Reverse primer: TTTCTTGCTGCCAGTCTGGAC
TATA box -amplicon:
CGGTTTGCTGCGGTAATCATGAGGATAAGAGAGCCACGAACCACGGCACTGATTTT
CAGTTCTGGGAAAATGGTGTGCACAGGAGCCAAGAGTGAAGAACAGTCCAGACTG
GCAGCAAGAAA
Ubiquitin (GenBank Accession No. BC000449) Ubiquitin Forward primer: ATTTGGGTCGCGGTTCTTG
Ubiquitin Reverse primer: TGCCTTGACATTCTCGATGGT
Ubiquitin C -amplicon:
ATTTGGGTCGCGGTTCTTGTTTGTGGATCGCTGTGATCGTCACTTGACAATGCAGAT
CTTCGTGAAGACTCTGACTGGTAAGACCATCACCCTCGAGG
TTGAGCCCAGTGACACCATCGAGAATGTCAAGGCA
SDHA (GenBank Accessio n No. NM 004168) SDHA Forward primer:
TGGGAACAAGAGGGCATCTG
SDHA Reverse primer: CCACCACTGCATCAAATTCATG
SDHA-amplicon TGGGAACAAGAGGGCATCTGCTAAAGTTTCAGATTCCATTTCTGCTCAGTATCCAGT
AGTGGATCATGAATTTGATGCAGTGGTGG
Oligonucleotide-based micro-array experiment protocol-Microarray fabrication Microarrays (chips) were printed by pin deposition using the MicroGrid II MGII
robot from BioRobotics Limited (Cambridge, UK). 50-mer oligonucleotides target sequences were designed by Compugen Ltd (Tel-Aviv, IL) as described by A. Shoshan et al, "Optical technologies and informatics", Proceedings of SPIE. Vol 4266, pp. 86-95 (2001). The designed oligonucleotides were synthesized and purified by desalting with the Sigma-Genosys system (The Woodlands, TX, US) and all of the oligonucleotides were joined to a C6 amino-modified linker at the 5' end, or being attached directly to CodeLink slides (Cat #25-6700-O1. Amersham Bioscience, Piscataway, NJ, US). The 50-mer oligonucleotides, funning the target sequences, were first suspended in Ultra-pure DDW (Cat # O1-866-lA Kibbutz Beit-Haemek, Israel) to a concentration of SOpM. Before printing the slides, the oligonucleotides were resuspended in 300mM sodium phosphate (pH 8.5) to final concentration of 150mM and printed at 35-40%
relative humidity at 21 °C.
Each slide contained a total of 9792 features in 32 subarrays. Of these features, 4224 features were sequences of interest according to the present invention and negative controls that were printed in duplicate. An additional 288 features (96 target sequences printed in triplicate) contained housekeeping genes from Human Evaluation Library2, Compugen Ltd, Israel.
Another 384 features are E.coli spikes 1-6, which are oligos to E Coli genes which are commercially available in the Array Control product (Array control- sense oligo spots, Ambion Inc. Austin, TX. Cat #1781, Lot #112K06).
Post-coupling processing of printed slides After the spotting of the oligonucleotides to the glass (CodeLink) slides, the slides were incubated for 24 hours in a sealed saturated NaCI humidification chamber (relative humidity 70-75%).
Slides were treated for blocking of the residual reactive groups by incubating them in blocking solution at 50°C for 15 minutes (lOml/slide of buffer containing O.1M Tris, SOmM
ethanolamine, 0.1% SDS). The slides were then rinsed twice with Ultra-pure DDW
(double distilled water). The slides were then washed with wash solution (lOml/slide.
4X SSC, 0.1%
SDS)) at 50°C for 30 minutes on the shaker. The slides were then rinsed twice with Ultra-pure DDW, followed by drying by centrifugation for 3 minutes at 800 rpm.
Next, in order to assist in automatic operation of the hybridization protocol, the slides were treated with Ventana Discovery hybridization station barcode adhesives.
The printed slides were loaded on a Bio-Optica (Milan, Italy) hematology staining device and were incubated for 10 minutes in SOmI of 3-Aminopropyl Triethoxysilane (Sigma A3648 lot #122K589). Excess fluid was dried and slides were then incubated for three hours in 20 mm/Hg in a dark vacuum desiccator (Pelco 2251, Ted Pella, Inc. Redding CA).
The following protocol was then followed with the Genisphere 900-RP (random primer), with mini elute columns on the Ventana Discovery HybStationT"'', to perform the microarray experiments. Briefly, the protocol was performed as described with regard to the instructions and information provided with the device itself. The protocol included cDNA
synthesis and labeling. cDNA concentration was measured with the TBS-380 (Turner Biosystems.
Sunnyvale, CA.) PicoFlour, which is used with the OliGreen ssDNA Quantitation reagent and kit.
Hybridization was performed with the Ventana Hybridization device, according to the provided protocols (Discovery Hybridization Station Tuscon AZ).
The slides were then scanned with GenePix 4000B dual laser scanner from Axon Instruments Inc, and analyzed by GenePix Pro 5.0 software.
Schematic summary of the oligonucleotide based microarray fabrication and the experimental flow is presented in Figures 4 and 5.
Briefly, as shown in Figure 4, DNA oligonucleotides at 25uM were deposited (printed) onto Amersham 'CodeLink' glass slides generating a well defined 'spot'. These slides are covered with a long-chain, hydrophilic polymer chemistry that creates an active 3-D surface that covalently binds the DNA oligonucleotides 5'-end via the C6-amine modification. This binding ensures that the full length of the DNA
oligonucleotides is available for hybridization to the cDNA and also allows lower background, high sensitivity and reproducibility.
Figure 5 shows a schematic method for performing the microarray experiments.
It should be noted that stages on the le$-hand or right-hand side may optionally be performed in any order, including in parallel, until stage 4 (hybridization). Briefly, on the left-hand side, the target oligonucleotides are being spotted on a glass microscope slide (although optionally other materials could be used) to form a spotted slide (stage 1). On the right hand side, control sample RNA and cancer sample RNA are Cy3 and Cy5 labeled, respectively (stage 2), to form labeled probes. It should be noted that the control and cancer samples come from corresponding tissues (for example, normal prostate tissue and cancerous prostate tissue).
Furthermore, the tissue from which the RNA was taken is indicated below in the specific examples of data for particular clusters, with regard to overexpression of an oligonucleotide from a "chip"
(microan-ay), as for example "prostate" for chips in which prostate cancerous tissue and normal tissue were tested as described above. In stage 3, the probes are mixed. In stage 4, hybridization is performed to form a processed slide. In stage 5, the slide is washed and scanned to form an image file, followed by data analysis in stage 6.
Cluster H61775 features 2 transcripts) and 6 segments) of interest, the names for which are given in Tables 1 and 2, respectively, the sequences themselves are given at the end of the application. The selected protein variants are given in table 3.
Table I - Transcripts of interest .Transcn .~ .;~-tName: .:~ 0~
'~ ~~ . ,~ SEQ-.~I
. ~ ,, D N
~p ~ ~--=.
..
-~~ ~:~~
=
H61775 T21 _ . ,, , . ..:
... .
Table 2 - Segments of interest --Segment ame ~-~~~ ~-SE ~ - ~ ~ T~:;~ w.~.~~
- ~ ~ ~ ~ ~ ~~r Q I D NO..
H61775 node 2 3 .:
~w ~ ~ ..~...~ .
~
H61775 node 4 H61775 node 6 H61775 node-8 6 H61775 node 0 7 H61775 node 5 g Table 3 - Proteins of interest otein -.
~ ~~ ~ ~ ~~
ame ~
~
. .
z ,.
~ , . ~,. , ;t, ' .'ts. ,...~ ~ ~n ., ix ~~ ~a. , ~'~ - ~, ~ ,~:'f~ z.~ ~~!1~~:.,.
~., ~~ H~
. a ,.,., .~x, 5 ~6 4 7 ~.;
~uh~..m~ , ~.a1 i..~ ~
~ :
~
H61775 P16 .,w .
..
~~ ~Lz. - N..~'.
Cluster H61775 can be used as a diagnostic marker according to overexpression of transcripts of this cluster in cancer. Expression of such transcripts in normal tissues is also given according to the previously described methods. The term "number" in the right hand column of the table and the numbers on the taxis of Figure 6 refer to weighted expression of ESTs in each category, as "parts per million" (ratio of the expression of ESTs for a particular cluster to the expression of all ESTs in that category, according to parts per million).
Overall, the following results were obtained as shown with regard to the histograms in Figure 6 and Table 4. This cluster is overexpressed (at least at a minimum level) in the following pathological conditions: brain malignant tumors and a mixture of malignant tumors from different tissues.
Table 4 - Normal tissue distribution Nam' o fTzssue _Number bladder 0 brain 0 colon 0 epithelial 10 general 3 breast g muscle 0 ovary 0 pancreas 0 prostate 0 uterus 0 Table S - P values and rat ios for expression in cancerous tissue r ~ u~. ~ ~~ P
r Name: of ~ ' ''= ~y -'~.~~SP2 R;4 Tissue. p f 2_~~x '~~ R3 .- ' ' SP1: ~ fi _ ek~.'I,~
~ % ' .._ . ., ~ ;. Y" ~ .",.
~rv ,.
~bladder~m ~3 E~.,S~c"..'firs . - , ..
R~
~O
. 3.8e-O13.2e-O12.5 4.6e-O1 1.9 e-l brain 8.8e-026.Se-021 3.5 4.1e-04 5.8 colon 5.6e-O16.4e-O11 1.1 1 1.1 epithelial 3.Oe-021.3e-O12.3e-022.1 3.2e-O1 1.2 general 1.3e-064.9e-OSl.Oe-076.3 l.Se-06 4.3 breast 4.7e-01 3.7e-O13.3e-O12.0 4.6e-O11.6 muscle 2.3e-O1 2.9e-O11.5e-O16.8 3.9e-O12.6 ovary 3.8e-O1 4.2e-O11.5e-012.4 2.6e-Ol1.9 pancreas 3.3e-O1 4.4e-Ol4.2e-O12.4 5.3e-O11.9 prostate 7.3e-O1 7.8e-OI6.7e-OI1.5 7.Se-O11.3 uterus 1.Oe-01 2.6e-O12.9e-O 2.6 5. I 1.8 l e-O1 As noted above, cluster H61775 Ieatures 2 transcript(s), which were listed in Table 1 above. A description of each variant protein according to the present invention is now provided.
Variant protein H61775 P16 according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) H61775 T21. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between H61775 P16 and Q9P2J2 (SEQ ID N0:953):
l.An isolated chimeric polypeptide encoding for H61775 P16, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 11 - 93 of Q9P2J2, which also corresponds to amino acids 1 - 83 of H61775 P16, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV corresponding to amino acids 84 - 152 of H61775 P16, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated polypeptide encoding for a tail of H61?75 P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV in H617?5 P16.
Comparison report between H61775 P16 and AAQ88495 (SEQ ID N0:954):
I .An isolated chimeric polypeptide encoding for I-161775 P 16, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 1 - 83 of AAQ88495, which also corresponds to amino acids 1 - 83 of H61775 P16, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the~sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV corresponding to amino acids 84 - 152 of H61775 P 16, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated polypeptide encoding for a tail of H61775 P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV in H61775 P16.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
secreted. The protein localization is believed to be secreted because both signal-peptide prediction programs predict that this protein has a signal peptide, and neither trans-membrane region prediction program predicts that this protein has a trans-membrane region..
Variant protein H61775 P16 also has the following norrsilent SNPs (Single Nucleotide Polymorphisms) as listed in Table 6, (given according to their positions) on the amino acid sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein H61775 P 16 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 6 - Amino acid mutations :. .,>cf ~~~.~'4~~w;?~~K~, n ~. Alternatue~ain~no~acid~ Pieviousl ;=known SNP 'ositiori s -Son:ayuno~acids 'i SNP?
p ~ ) ~ b - ( ) Y
;~. . , ., --~,:..E ~ ~ ~ ~ ~
. , ~r~ : <
~
~
~se uence ~: ~. , ~ .~~
, ~~- .: ~ ~ . v;
_ .. ~
zq P. ,~ -~~ ~ ~ . ~~ ., : . .~ ..
~ ~ . ~ ~ ~ z , ~"~.
~. t-x~ .~ _..~.~ ~_. . ,_ ~~ ___~ N~ , ~ .w .. ...__ 14 I -> T No 138 G -> R No 34 G -> E Yes 48 G -> R No 91 R -> * Yes Variant protein H61775 P16 is encoded by the following transcript(s): H61775 T21, for which the sequences) is/are given at the end of the application. The coding portion of transcript H61775 T21 is shown in bold; this coding portion starts at position 261 and ends at position 716. The transcript also has the following SNPs as listed in Table 7 (given according to their position on the nucleotide sequence, with the alternative nucleic acid listed;
the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein H61775 P16 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 7 - Nucleic acid SNPs a., :PfY' satFwv'.,.~.. . ":..~:rn .yd~, . ..f.:. '~, .iff0.,yx., "~ ~,,x ~..-. y : _ a , . : ,m ~i '.
z. nucleotide ~'. ~;Altemat ~ uel . e- acidp - ~ .ev ou 1~ .known. SNP
S ~,pas~han ova .. ~ e~..; ~n ~ ' o y , TTws -._ ' ' ~ ~,, ~F,,.,.w .' sx'."' :e:~.cai. z ~.,re ,. x,. 'fi" '~ arY, ,i m,..,.r ~~,"sm,~
equ~nc~ .. ~ ~ ~. ate. - ~ ~ r- .: ~s " ,. ~.
" ,"i~.",~, .." w.='~.. ,..,f'. x.r~' ..~~,4:::..a.. .~.r~,_ : ~R~. ~,: m~<., ~ ~~cr;, °a . .
117 T -> C Yes 200 T -> C No 672 G -> C No 222 T -> C Yes 301 T -> C No 361 G -> A Yes 377 G -> A No 400 -> C No 402 G -> C No 531 C -> T Yes 566 ~ T -> C ~ No Variant protein I-161775 P17 according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) H61775 T22. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between H61775 P17 and Q9P2J2:
l.An isolated chimeric polypeptide encoding for H61775 P17, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 11 - 93 of Q9P2J2, which also corresponds to amino acids 1 - 83 of H61775 P17.
Comparison report between H61775 P17 and AAQ88495:
l.An isolated chimeric polypeptide encoding for H61775 P17, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 1 - 83 of AAQ88495, which also corresponds to amino acids 1 - 83 of H61775 P17.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
secreted. The protein localization is believed to be secreted because both signal-peptide prediction programs predict that this protein has a signal peptide, and neither trans-membrane region prediction program predicts that this protein has a trans-membrane region..
Variant protein H61775 P17 also has the following norrsilent SNPs (Single Nucleotide Polymorphisms) as listed in Table 8, (given according to their positions) on the amino acid sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein H61775 P17 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 8 - Amino acid mutations 3 7k"' '::' RE- -r ...c '.,>: _ r ~.".- - '~'...
'.' ~~~~?. -~. . , 'n _,.. T~:~e"~"
.,..2 3v'..~ri c~ ,k9~ f" ~"w .' .. vy~-35".
7 ° '.:. ~5 ..,..h~ ., .3.,. ,:.~ ,..~a' , , ~,~, .~.t,' ..;t" n ,.l.y- ~.".o. .; .~;, M,..,.,. .. 2 . Y~e.~Fy: -~
~4 ' , ' ,: % ' "..i,'", ... 'GSS -.y . x. dl v:~F°a~~_..
osati n .s ~on~ammo~acid F..Alternatiue~ammo.,acid s= m I'xemousl ~'known,-SNP
. x".
,k..h'.'a~w, ". "'& ~ 'f'°.. r w~'~:3F
R ". ~ -:.. ,."~.Xt' .. ~....3~°' ~~ ~.:~~ ~ ~z '-2~Y~W~~ -.:,W~ -. 'SYZ" W: ~. n~..-- __.",w I -.;~ rte.
'~RL ' ">~~~~ ', ~ ~ , , ~,.,'3 f.: .~ m~ s,,w Y'". Fi : ~, % aFSf ."-..,~, .. , t~~~ , V ~ -, s L.. S::":, " ~ . ., 5 ~s'' ,~Y . , S.f. ' ,,~y"~'T-.~,.
'K
_~;,x2 I .:.>~s:.:", ,:b~
-~se uencer. ~,~T_,. "
,..,y. ' -~'. .,'-.. S~m,. , ,z sr,~ ~x...~,~..,..:'~... terse. ~a', ,.._r ~.-~-:~- .~.~ ~, Wig...-.~,,~.: v- '~. ,;~::~y:~;.- ~,-<..
..,d~" ~..., " ~ ' "~Y,...,.,.3"t'.r N .,~~ ,... ,. x'=~,>w.~~,.;.,"
14 "xI ->" T " No 34 G -> E Yes 48 G -> R No Variant protein H61775 P17 is encoded by the following transcript(s): H61775 T22, for which the sequences) is/are given at the end of the application. The coding portion of transcript H61775 T22 is shown in bold; this coding portion starts at position 261 and ends at position 509. The transcript also has the following SNPs as listed in Table 9 (given according to their position on the nucleotide sequence, with the alternative nucleic acid listed;
the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein H61775 P17 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 9 - Nucleic acid SNPs ~ :w,~ ~ ~ - ~ ~ r ~_~~"F,.,.< . ... :° ~:; !~
~.'F
r "1 ~ id .;, :eYi = : 1 ,s NPR ~ osiiwton on.nu~l"eatrde , _ Iteriiatxve~nuc a c c ~ 9 x out. . _I~no~ S
,b ~ . a =.sequence F ~ ~ ,~ a.~ w ~~ .
.,_ _ .,,>W ~-.,.. ~~ r.,. ~ w;
117 T -> C ~ Yes 200 T -> C No 222 T -> C Yes 301 T -> C No 361 G -> A Yes 377 G -> A No 400 -> C No 402 G -> C No 596 T -> A Yes Ac nc,ted above. cluster61775 features fi H se~ment(sl. which were listed in Table 2 above and for which the sequences) are given at the end of the application. These segments) are portions of nucleic acid sequences) which are described herein separately because they are of particular interest. A description of each segment according to the present invention is now provided.
Segment cluster H61775 node 2 according to the present invention is supported by 17 libraries. The number of libraries was determined as previously described.
This segment can be found in the following transcript(s): H61775 T21 and H61775 T22. Table 10 below describes the starting and ending position of this segment on each transcript.
Table 10 - Segment location on transcripts 'S ~~~;,.at ~a~~ ~"'~~'.. _ ~*. a : y = s'.
,nyaPH..,.,.,.u. .. ~t~ -.. .~, ..~y , 5'~~, ~~ y.. ~Se . n end~n asz~on s a . ent~ last n . g : tt, a ~g ~Traaascn ,.name ~' . osxlran- .~ ~~iro, ~ ,5 gm.~ ~. : ~....~_~a'.
~ ~.._.,. ~
;a~M ~., rya =5 ~~'"~;ar ' ~ - ~
bY '-- ~
~ '-sF s , .. ..
.~ .. .. .....~ .
m , , ,. ..: ,4u;a. .
",rz~. ...~,.,~s... ~ c .
.._ , ,fir ...- "a.3:.: ~, ~:dr ~' -~.""a. .
s .~:'~ ;.. r- S~e. ';- ,?~
-~' ~ , . ":.s~.
s ~-. ,~~,.' "~ .o.
~ ~" e ' ~'z~~ - x : -:~;'~
s~ ~
, ; : ,.
, . .,. .
a ,. .
. .. .
, .. x_ ...
. ..
Segment cluster H61775 node 4 according to the present invention is supported by 20 libraries. The number of libraries was determined as previously described.
This segment can be found in the following transcript(s): H61775 T21 and H61775 T22. Table 11 below describes the starting and ending position of this segment on each transcript.
Table 11 - Segment location on transcripts a,,. , ~ ~:~~ ~w v '~", ~Transe 't~riame;~ Se yen ~star=tin Se enl endln ~ ' osytton ,-~~ , . Lyon . h ~;t . _. ~ -~'I?~
~?p ~ ~ :~
~B P: ~ p _a -~
~ ~ ~~~ r ,,._~~
y ~-. _ _ ~. ~ : m_ .
>_ ~ ~, .. ~;,.
: ~ _~: ~ f. : ~ry - ~ ~ s ~ ' ~o :
~:~_: ~ ~~ ~s Segment cluster H61775 node 6 according to the present invention is supported by 1 libraries. The number of libraries was determined as previously described.
This segment can be found in the following transcript(s): H61775 T22. Table l2 below describes the starting and ending position of this segment on each transcript.
Table 12 - Segment location on transcripts ,~~ ~. ~ ~ ~ -X R J w .F. ~&. 'w~ ~I. , Transcn t<~iiame -~ ~~ " ent startnri S.e mrntwendm ~ .osvtson M. ositaon .W a..~
~ g p ~? . g g~l? ~ 5 ~
, , ., s ~ ~. ~
. .
~~
~
Segment cluster H61775 node 8 according to the present invention is supported by 5 libraries. The number of libraries was determined as previously described.
This segment can be found in the following transcript(s): H61775 T21. Table 13 below describes the starting and ending position of this segment on each transcript.
Table 13 - Segment location on transcripts rte: ~~- .. ~ .~ -~~;
~ -:~ - ~, .e. t ,. .x , ltl _ ~ _ ti ~a_ z ~p ~ n ..~. ~ - a _nt~a~dan ~ os~
- x. . ~e ent s ~z~t~ _ n ~~~~
Tra.~s ~ t,.name;: os a . g I?
. b= ;~~ v: ,. .
~ - :.
~ ~
~ na ~
H a , ~
~ ~ , ..~, .w.
~..~ . ~,: ~ ~
-~ ' .. ~ ~ ~ ~ f~ , "~ ' ~ .,r:
n~ ~
~~.:: ~;
. ~~. s _' ' ' -to :r.., S _b ~ , ..zm".~,.,.,. ~z "~ ~~
~ , - u__~.~.:
- Y ,re'.%.
According to an optional embodiment of the present invention, short segments related to the above cluster are also provided. These segments are up to about 120 by in length, and so are included in a separate description.
Segment cluster H61775 node 0 according to the present invention is supported by 4 libraries. The number of libraries was determined as previously described.
This segment can be found in the following transcript(s): H61775 T21 and H61775 T22. Table 14 below describes the starting and ending position of this segment on each transcript.
Table 14 - Segment location on transcripts H61775T2l I 86 Segment cluster H61775 node 5 according to the present invention can be found in the following transcript(s): H61775 T22. Table 15 below describes the starting and ending position of this segment on each transcript.
Table I S - Segment location on transcripts ~..1'.z2., ~2 .,rn.2,~- .~.d; ~. ~ .-~3r3.'~:-.wL:~-..~....:
'~-. ~ ~ . ,~ .....-3~,~" w"~,.
'T~. ,.~..>.~Yx ... - ~ '.~- i~F'x .. "..R
4.~'vr '~f" , ta~' Se' . entstartm ~ ..~Se ent endxn o~xtlon Transcn . t:~name~: osxtion,~; _, .:; ..y . ~~ , ~ .g P ~_ ~ p ~,~ ~? ~
~ ' ~
~__ .~. _. .7 a .,.~ -Fns ; :.~ ~"~~ ~, ~~ ..v~
' ,." ,.
.'.. ~ ~.--,~ .~, ~
-.n..~ ~_,. ~ - ~
~. : w -W
~~ ~a 6.~, ;~:.e5.
~_~. ty'-.- ~-:.,.,'.
:~'..,~~~5'" , ..,.. ..: w~%T ',~.CS
. L'.~'.~ ..,.s Variant protein alignment to the previously known protein:
Sequence name: /tmp/PswORJLCti/aLAXQjXh07:Q9P2J2 Sequence documentation:
Alignment of: H61775-P16 x Q9P2J2 ..
Alignment segment 1/1:
Quality: 803.00 Escore: 0 Matching length: 83 Total length: 83 Matching Percent Similarity: 100.00 Matching Percent Identity: 100.00 Total Percent Similarity: 100.00 Total Percent Identity: 100.00 Gaps: 0 Alignment:
IIIIIIIIIIIIIIIIillllllllllllllll 20 Sequence name: /tmp/PswORJLCti/aLAXQjXh07:AAQ88495 Sequence documentation:
Alignment of: H61775 P16 x AAQ88495 ..
Alignment segment 1/1:
Quality: 803.00 Escore: 0 Matching length: 83 Total length: 83 Matching Percent Similarity: 100.00 Matching Percent Identity: 100.00 Total Percent Similarity: 100.00 Total Percent Identity: 100.00 S Gaps: 0 Alignment:
IIIIiillllllllllllllllllllllllllllllllllllllilllll IIIIIIIIIIIIIIIIIIIIIIIiillllllll Sequence name: /tmp/naab8yR3GC/pSM412IL5o:Q9P2J2 Sequence documentation:
Alignment of: H61775 P17 x Q9P2J2 , .
Alignment segment 1/1:
Quality: 803.00 Escore: 0 Matching length: 83 Total length: 83 Matching Percent Similarity: 100.00 Matching Percent Identity: 100.00 Total Percent Similarity: 100.00 . Total Percent Identity: 100.00 Gaps: 0 Alignment:
IIIIIIIIIIIIIIIIilllllllllillllllilllllllillllllll IIIIIIIIIIIIIIIIIIilllllillllllli Sequence name: /tmp/naab8yR3GC/pSM412IL5o:AAQ88495 Sequence documentation:
Alignment of: H61775_P17 x AAQ88495 ..
Alignment segment 1/1:
Quality: 803.00 Escore: 0 Matching length: 83 Total length: 83 Matching Percent Similarity: 100.00 Matching Percent Identity: 100.00 Total Percent Similarity: 100.00 Total Percent Identity: 100.00 Gaps: 0 Alignment:
IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
IIIIIIIIIIIIIIIIIIIIIIillilllllll 25 Expression of immunoglobulin superfamily, member 9 H61775 transcripts which are detectable by amplicon as depicted in sequence name H61775seg8 in normal and cancerous ovary tissues.
Expression of immunoglobulin superfamily, member 9 transcripts detectable by or according to H61775seg8, H61775seg8 amplicon(s) and H61775seg8F2 and H61775seg8R2 primers was measured by real time PCR. In parallel the expression of four housekeeping genes:
PBGD (GenBank Accession No. BC019323; amplicon - PBGD-amplicon), HPRT1 (GenBank Accession No. NM 000194; amplicon - I-IPRT1-amplicon), and SDHA (GenBank Accession No. NM 004168; amplicon - SDHA-amplicon), GAPDI-I (GenBank Accession No.
BC026907;
GAPDI-1 amplicon) was measured similarly. For each RT sample, the expression of the above amplicon was normalized to the geometric mean of the quantities of the housekeeping genes.
The normalized quantity of each RT sample was then divided by the median of the quantities of the normal post-mortem (PM) samples (Sample Nos. 45-48,71, Table l, "Tissue samples in testing panel"), to obtain a value of fold up-regulation for each sample relative to median of the normal PM samples.
Figure 7 is a histogram showing over expression of the above-indicated immunoglobulin superfamily, member 9 transcripts in cancerous ovary samples relative to the normal samples.
(Values represent the average of duplicate experiments. Error bars indicate the minimal and maximal values obtained As is evident from Figure 7, the expression of immunoglobulin superfamily, member 9 transcripts detectable by the above amplicon(s) in cancer samples was significantly higher than in the norrcancerous samples (Sample Nos. 45-48, ,71 Table 1, "Tissue samples in testing panel") and including benign samples (samples No. 56, 62, 64). Notably an over-expression of at least 5 fold was found in 21 out of 43 adenocarcinoma samples.
Statistical analysis was applied to verify the significance of these results, as described below.
The P value for the difference in the expression levels of immunoglobulin superfamily, member 9 transcripts detectable by the above amplicon(s) in ovary cancer samples versus the normal tissue samples was determined by T test as 2.76E-4.
The above value demonstrates statistical significance of the results.
Primer pairs are also optionally and preferably encompassed within the present invention; for example, for the above experiment, the following primer pair was used as a norr limiting illustrative example only of a suitable primer pair: H61775seg8F2 forward primer; and H61775seg8R2 reverse primer.
The present invention also preferably encompasses any amplicon obtained through the use of any suitable primer pair; for example, for the above experiment, the following amplicon was obtained as a norrlimiting illustrative example only of a suitable amplicon: H61775seg8 H61775seg8F2 (SEQ ID N0:955) GAAGGCTCTTGTCACTTACTAGCCAT
H61775seg8R2 (SEQ ID N0:956) TGTCACCATATTTAATCCTCCCAA
Amplicon (SEQ ID N0:957) GAAGGCTCTTGTCACTTACTAGCCATGTGATTTTGGAAAGAAACTTAACATTAATTC
CTTCAGCTACAATGGAATTCTTGGGAGGATTAAATATGGTGACA
Expression of immunoglobulin superfamily, member 9 H61775 transcripts which are detectable by amplicon as depicted in sequence name H61775seg8 in different normal tissues.
Expression of immunoglobulin superfamily, member 9 transcripts detectable by or according to H61775 seg8 amplicon(s) and H61775 segBF and H61775 segBR was measured by real time PCR. In parallel the expression of four housekeeping genes -RPL19 (GenBank Accession No. NM 000981; RPL19 amplicon), TATA box (GenBank Accession No.
NM-003194; TATA amplicon), Ubiquitin (GenBank Accession No. BC000449; amplicon -Ubiquitin-amplicon) and SDHA (GenBank Accession No. NM 004168; amplicon - SDHA-amplicon) was measured similarly. For each RT sample, the expression of the above amplicon was normalized to the geometric mean of the quantities of the housekeeping genes. The normalized quantity of each RT sample was then divided by the median of the quantities of the ovary samples (Sample Nos. 18-20, Table 2 "Tissue samples in normal panel", above), to obtain a value of relative expression of each sample relative to median of the ovary samples.
The results are described in Figure 8, presenting the histogram showing the expression of H61775 transcripts which are detectable by amplicon as depicted in sequence name H61775seg8, in different normal tissues. Amplicon and primers are as above.
DESCRIPTION FOR CLUSTER HSAPHOL
Cluster HSAPHOL features 7 transcripts) and 18 segments) of interest, the names for which are given in Tables 1 and 2, respectively, the sequences themselves are given at the end of the application. The selected protein variants are given in table 3.
Table I - Transcripts of interest _~ ~ ~w, fw~~~ ~Cw.... ~ ~~, ~.'~rW.~,~_ ~"~.'v, T~ranscn txN me'~ ~ '~ .~ ...:',2'n' '&~"" p4~~'%..
_' ' ~--W ~SE ~ ~ID'~ O~' a ~ ,, ~~
s- - ~~. - ~ _ _ ~
~ ~ s s~'r ~.s"~' ~
T ra,~ ..a ~ ,."~~,a" " f~ '~~. .
~"~ ~. ,~$;r~ _ ~ . ~Y'r'9~~~'s,s~, , n a . , .-a k ~
Table 2 - Segments of interest .y-.,-. :z- .~; ~~ _ .' .C.. ~ -,~~ Rk ,.-a Y
Se~ a t'~T e~~. ~ ~:~' ~; ASE ~ ~II3x~NO~~ ~, ~ .>.
~ & ~ ~:~ ~x '.,~ x . w "'"<. kF . .~ 3 . ~b~ G~ : 7. "~x ~. i'r 7 _, ,' ~:1. 4,or i , ~:.'. .- ; .. ' 'u' ~ a. s ,~ , -4 ~ y ~~s.
~ ." ,A~~,.~ui- '4 HSAPHOL node 11 18 HSAPHOL node 13 19 HSAPHOL node 15 20 HSAPI-IOL node 19 21 HSAPI-IOL node 2 22 HSAPHOL node 21 23 HSAPI-IOL node 23 24 HSAPHOL node 26 25 HSAPI-IOL node 28 26 HSAPHOL node 38 27 HSAPHOL node 40 28 HSAPHOL node 42 29 HSAPHOL node 16 30 HSAPHOL node 25 31 HSAPHOL node 34 32 HSAPHOL node 35 33 HSAPHOL node 36 34 HSAPHOL node 41 ~ S
Table 3 - Proteins of interest ~roteua a~. ~ n~~Q ~D~~t~ .~, ~ ~~, ~,~
~r~ ~~ ~ a . ~m ''r~x s~ ~.+: a ~ -A
These sequences are variants of the known protein Alkaline phosphatase, tissue-nonspecific isozyme precursor (SwissProt accession identifier PPBT,HUMAN;
known also according to the synonyms EC 3.1.3.1; AP-TNAP; Liver/bone/kidney isozyme;
TNSALP), SEQ
ID NO: 36, referred to herein as the previously known protein.
The variant proteins according to the present invention are variants) of a known diagnostic marker, called Alkaline Phosphatase.
Protein Alkaline phosphatase, tissue-nonspecific isozyme precursor is known or believed to have the following funetion(s): THIS ISOZYME MAY PLAY A ROLE IN SKELETAL
MINERALIZATION. The sequence for protein Alkaline phosphatase, tissue-nonspecific isozyme precursor is given at the end of the application, as "Alkaline phosphatase, tissue-nonspecific isozyme precursor amino acid sequence". Known polymorphisms for this sequence are as shown in Table 4.
Table 4 - Amino acid mutations for Known Protein , SNPp,~osation(s)rori Comment ~».:~~ ~' .,,~~ ~, ima acid a nce . ~~~ ~ ~~ ~;~.
_.~_,t-1-. ~ >r. .~.~.;_:..,. ..~: -~.."~".....~',"w >s, ~..,w. ~;~ ,.
...~:.',-~, . : , ,,: .~a> ~u_ a.a ':~.w n".~....em~.,. '.. Via'':.'~.
28 Y -> C (in hypophosphatasia; infantile; 7% of activity).
/FTId=VAR 013972.
33 A -> V (in hypophosphatasia). /FTId=VAR 006147.
111 A -> T (in hypophosphatasia; odonto).
/FTId=VAR 006151.
116 A -> T (in hypophosphatasia; loss of activity).
/FTId=VAR 013977.
120 G -> R (in hypophosphatasia). /FTId=VAR 013978.
129 G -> R (in hypophosphatasia). /FTId=VAR 013979.
132 A -> V (in hypophosphatasia). /FTId=VAR 013146.
134 T -> N (in hypophosphatasia; 9% of activity).
/FTId=VAR 011082.
136 R -> H (in hypophosphatasia; moderate; 33% of activity).
/FTId=VAR 006152.
152 R -> H (in hypophosphatasia). /FTId=VAR O 13980.
162 G -> V (in hypophosphatasia; severe; 1 % of activity).
/FTId=VAR 006153.
170 N -> D (in hypophosphatasia). /FTId=VAR
013981.
40 A -> V (in hypophosphatasia; 2% of activity).
/FTId=VAR 011081.
171 H -> Y (in hypophosphatasia; severe;
2% of activity).
/FTId=VAR 006154.
I 76 A -> T (in hypophosphatasia). /FTId=VAR
0 t 1083.
177 A -> T (in hypophosphatasia; adult type).
/FTId=VAR 006155.
179 A -> T (in hypophosphatasia). /FTId=VAR
006156.
181 S -> L (in hypophosphatasia; 1 % OF
activity).
lFTId=VAR 013982.
184 R -> W (in hypophosphatasia; loss of activity).
/FTId=VAR 013983.
191 E -> G (in hypophosphatasia; odonto).
/FTId=VAR 006157.
191 E -> K (in hypophosphatasia; moderate;
frequent mutation in European countries). /FTId=VAR 006158.
201 C -> Y (in hypophosphatasia). /FTId=VAR
006159.
207 Q -> P (in hypophosphatasia). /FTId=VAR
006160.
51 A -> V (in hypophosphatasia). /FTId=VAR
013973.
211 N -> D (in hypophosphatasia). /FTId=VAR
013984.
220 G -> V (in hypophosphatasia; odonto).
/FTId=VAR 013985.
223 R -> W (in hypophosphatasia; 3% of activity).
/FTId=VAR 013986.
224 K -> E (in hypophosphatasia; infantile;
partial loss of activity). /FTId=VAR 011084.
235 E -> G (in hypophosphatasia). /FTId=VAR
013987.
246 R -> S (in hypophosphatasia; 4% of activity).
/FTId=VAR 011085.
249 G -> V (in hypophosphatasia; partial loss of activity).
IFTId=VAR 013988.
263 H -> Y (common polymorphism). lFTld=VAR
006161.
289 L -> F (in hypophosphatasia). /FTId=VAR
006162.
291 E -> K (in hypophosphatasia; moderate;
8% of activity).
/FTId=VAR 013989.
62 M -> L (in hypophosphatasia; moderate;
27% of activity).
/FTId=VAR 006148.
294 D -> A (in hypophosphatasia). /FTId=VAR
006163.
294 D -> Y (in hypophosphatasia). /FTId=VAR
013990.
306 D -> V (in hypophosphatasia). /FTId=VAR
006164.
326 G -> R (in hypophosphatasia; in a patient carrying also lys-291). /FTId=VAR 013991.
327 F -> G (in hypophosphatasia; requires 2 nucleotides substitutions). /FTId=VAR 013992.
327 F -> L (in hypophosphatasia; childhood).
/FTId=VAR 006165.
334 G -> D (in hypophosphatasia). /FTId=VAR
006166.
348 A -> T (in hypophosphatasia). /FTId=VAR
011086.
378 D -> V (in hypophosphatasia; loss of activity).
/FTId=VAR 006167.
381 H -> R (in hypophosphatasia). /FTId=VAR
O 11087.
63 G -> V (in hypophosphatasia; loss of activity).
IFTId=VAR 013974.
382 V -> I (in hypophosphatasia). IFTId=VAR
006168.
391 R -> C (in hypophosphatasia; moderate;
10% of activity).
/FTId=VAR 013993.
399 A -> S (in hypophosphatasia). IFTId=VAR
013994.
406 D -> G (in hypophosphatasia; 15% of activity).
/FTId=VAR Ol 1088.
423 V -> A (in hypophosphatasia; l6% of activity).
/FTId=VAR 013995.
426 G -> C (in hypophosphatasia; infantile;
partial loss of activity).IFTId=VAR 011089.
436 Y -> H (in hypophosphatasia). IFTId=VAR
006169.
445 S -> P (in hypophosphatasia; severe;
2% of activity).
lFTId=VAR 013996.
450 R -> C (in hypophosphatasia; severe;
4% of activity).
/FTId=VAR 013997.
450 R -> H (in hypophosphatasia). /FTId=VAR
011090.
71 R -> C (in hypophosphatasia). /FTId=VAR
006149.
456 G -> R (in hypophosphatasia; loss of activity).
/FTId=VAR 011091.
459 V -> M (in hypophosphatasia; infantile).
/FTId=VAR 013998.
473 G -> S (in hypophosphatasia). /FTId=VAR
013999.
476 E -> K (in hypophosphatasia). /FTId=VAR
006170.
478 N -> I (in hypophosphatasia; 9% of activity).
/FTId=VAR 011092.
489 C -> S (in hypophosphatasia; 9% of activity).
/FTId=VAR 011093.
490 I -> F (in hypophosphatasia; odonto;
partial loss of activity). /FTId=VAR 014000.
491 G -> R (in hypophosphatasia). IFTId=VAR
522 V -> A. iFTId=VAR 011094.
29 W -> A
71 R -> H (in hypophosphatasia). /FTId=VAR
104 N -> K
71 R -> P (in hypophosphatasia). /FTId=VAR
006150.
75 I G -> S (in hypophosphatasia; severe; 3.5% of activity).
/FTId=VAR 013976.
Protein Alkaline phosphatase, tissue-nonspecific isozyme precursor localization is believed to be attached to the membrane by a GPI-anchor.
The following GO Annotations) apply to the previously known protein. The following annotations) were found: skeletal development; ossification; metabolism, which are annotations) related to Biological Process; magnesium binding; alkaline phosphatase;
hydrolase, which are annotations) related to Molecular Function; and integral membrane protein, which are annotations) related to Cellular Component.
The GO assignment relies on information from one or more of the SwissProt/TremBl Protein knowledgebase, available from <http://www.expasy.ch/sprot/>; or Locuslink, available from <http://www.ncbi.nlm.nih.gov/projects/LocusLink/>.
As noted above, cluster HSAPHOL features 7 transcript(s), which were listed in Table 1 above. These transcripts) encode for proteins) which are variants) of protein Alkaline phosphatase, tissue-nonspecific isozyme precursor. A description of each variant protein according to the present invention is now provided.
Variant protein HSAPHOL P2 according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) HSAPHOL T4. An alignment is given to the known protein (Alkaline phosphatase, tissue-nonspecific isozyme precursor) at the end of the application. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between HSAPHOL P2 and AAH21289 (SEQ ID NO: 36):
l.An isolated chimeric polypeptide encoding for HSAPHOL P2, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PHSGPAAAFIRRRGWWPGPRCA corresponding to amino acids 1 - 22 of HSAPI-iOL P2, second amino acid sequence being at least 90 % homologous to PATPRPLSWLRAPTRLCLDGPSPVLCA corresponding to amino acids 1 - 27 of AAH21289, which also corresponds to amino acids 23 - 49 of HSAPHOL P2, and a third amino acid sequence being at least 90 % homologous to EKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFLGDGMGVSTVTAARILKGQL
HHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLCGVKANEGTVGVSAAT
MPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDVEYESDEKARGTRLD
GLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEPGDMQYELNRNNVT
DPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALHEAVEMDRAIGQAG
SLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYK
VVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQN
YVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 83 - 586 of AAH21289, which also corresponds to amino acids 50 -553 of HSAPHOL P2, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
2.An isolated polypeptide encoding for a head of HSAPHOL P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence PHSGPAAAFIRRRGWWPGPRCA of HSAPHOL P2.
3.An isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P2, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise AE, having a structure as follows: a sequence starting from any of amino acid numbers 49-x to 49; and ending at any of amino acid numbers SO+ ((n-2) - x), in which x varies from 0 to rr2.
Comparison report between HSAPHOL P2 and PPBT HUMAN
1.An isolated chimeric polypeptide encoding for HSAPHOL-P2, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PHSGPAAAFIRRRGWWPGPRCAPATPRPLSWLRAPTRLCLDGPSPVLCA
corresponding to amino acids 1 - 49 of HSAPHOL P2, second amino acid sequence being at least 90 % homolo~us to EKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFLGDGMGVSTVTAARILKGQL
HHNFGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLCGVKANEGTVGVSAAT
ERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAAYAHSADRDWYSDNE
MPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDVEYESDEKARGTRLD
GLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEPGDMQYELNRNNVT
DPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALHEAVEMDRAIGQAG
SLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYK
VVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQN
YVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 21 - 524 of PPBT HUMAN, which also corresponds to amino acids 50 -553 of HSAPHOL P2, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
2.An isolated polypeptide encoding for a head of HSAPHOL P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence PHSGPAAAFIRRRGWWPGPRCAPATPRPLSWLRAPTRLCLDGPSPVLCA of HSAPHOL P2.
3.An isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P2, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about SO amino acids in length, wherein at least two amino acids comprise AE, having a structure as follows: a sequence starting from any of amino acid numbers 49-x to 49; and ending at any of amino acid numbers 50+ ((rr2) - x), in which x varies from 0 to rr2.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
membrane. The protein localization is believed to be membrane because although it is a partial protein, because both trans-membrane region prediction programs predict that this protein has a trans-membrane region, and similarity to known proteins suggests a GPI
anchor.Variant protein HSAPHOL P2 also has the following non-silent SNPs (Single Nucleotide Polymorphisms) as listed in Table 5, (given according to their positions) on the amino acid sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL P2 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 5 -Amino acid mutations a- "., "~~.F~. v~'X.. ,: . ,1' " ,. iT'~
~~ Y ~~. ., ,ES ~ W ' a ' non s : n-ammo° cid-;- Alte - atme~ mo. am . Preu~ousl known S .; "
SNP o~szt ; ~(. o , a gin, am d(s)." -; P~ y~ : ,~.
z ~a ~' ~ x~~ s~s~.~.' , 'i. -y "°,~,si~~a. a ,;icY & ". . t r i.., ~f F. Y
-. ~~,..'s. ~"a " c w:s~F.,~. ~t ~,~'r' .sr~m &a...
153 N -> S Yes 172 Q -> No 551 V -> A No 206 A -> No 272 R -> No 292 Y -> H Yes 342 V -> No 344 V -> No 354 K -> No 354 K -> Q No 380 E -> No Variant protein HSAPHOL P2 is encoded by the following transcript(s): HSAPHOL
T4, for which the sequences) is/are given at the end of the application. The coding portion of transcript HSAPHOL T4 is shown in bold; this coding portion starts at position 1 and ends at position 1659. The transcript also has the following SNPs as listed in Table 6 (given according to their position on the nucleotide sequence, with the alternative nucleic acid listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL P2 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 6 - Nucleic acid SNPs ;~.: ~ r _. ~",.,~ ,. . ~ :.==.
~~ ~-_ -~~ ~Previousl"' ~ kno n-SNP? . .. . ~.
SNP os~frop on nucleotide : ::Alternarivegnucleic acid~~. w _ ~.~p ,r'',~~r. a~~. ,:i >a. ~ »~a~-"=..,Fz ..~'r~ t~a°-~...~~.°
~~a'"'.,..'c~.r'''~'"~' ~r~~rri =...~..~-.-c ..~~.",~ ,.°. °cx.., iwm~.,~.-_..,:..~~;a.n~-~~~y ~...-.~'ss._. .y° a...,~ ~x ,_... ~..~F,-x:.~a.
417 C -> T Yes 458 A -> G Yes 1140 G -> No 1509 C -> T Yes 1629 G -> T Yes 1652 T -> C No 1727 C -> T Yes 1788 G -> A Yes 1895 A -> C Yes 2050 C -> T Yes 2095 A -> G Yes 2240 G -> No 516 G -> No 2347 -> A No 2364 T -> G No 617 C -> No 815 G -> No 874 T -> C Yes 1026 G -> No 1032 G -> No 1060 A -> No 1060 A -> C No Variant protein HSAPHOL P3 according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) HS APHOL T5. An alignment is given to the known protein (Alkaline phosphatase, tissue-nonspecific isozyme precursor) at the end of the application. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between HSAPHOL P3 and AAH21289:
1.An isolated chimeric polypeptide encoding for HSAPHOL P3, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVP
corresponding to amino acids 63 - 82 of AAH21289, which also corresponds to amino acids 1 -20 of HSAPHOL P3, and a second amino acid sequence being at least 90 % homologous to GMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYL
CGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSA
AYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTD
VEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFE
PGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQAL
HEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKK
PFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKG
PMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSV
LF corresponding to amino acids 123 - 586 of AAH21289, which also corresponds to amino acids 21 - 484 of HSAPHOL P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P3, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise PG, having a structure as follows: a sequence starting from any of amino acid numbers 20-x to 20; and ending at any of amino acid numbers 21+ ((m2) - x), in which x varies from 0 to n-2.
Comparison report between I-ISAPI-IOL P3 and PPBT HUMAN
l .An isolated chimeric polypeptide encoding for HSAPHOL P3, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVP
corresponding to amino acids 1 - 20 of PPBT HUMAN, which also corresponds to amino acids 1 -20 of 1-ISAPI-IOL_P3, and a second amino acid sequence being at least 90 %
homologous to GMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYL
CGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSA
AYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTD
VEYESDE.KARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFE
PGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQAL
HEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKK
PFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKG
PMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSV
LF corresponding to amino acids 61 - 524 of PPBT_HUMAN, which also corresponds to amino acids 21 - 484 of HSAPHOL P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P3, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise PG, having a structure as follows: a sequence starting from any of amino acid numbers 20-x to 20; and ending at any of amino acid numbers 21+ ((n-2) - x), in which x varies from 0 to rr2.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
membrane. The protein localization is believed to be membrane because of manual inspection of known protein localization and/or gene structure, and/or similarity to known proteins..
Variant protein HSAPHOL P3 also has the following non-silent SNPs (Single Nucleotide Polymorphisms) as listed in Table 7, (given according to the it positions) on the amino acid sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL P3 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 7 - Amino acid mzrtatiorrs ~,> :- ~s~:~..
. ~.. ,h~u.- . ~"~.~,. ..
t m. s, ax.r. .r id '~Alteriiatiue .,.-. ~ . , w~ .
SNI'= ositton s on~amtno ac 1? ~, ( ) ~ r am no,.aqid~.s- ~~'~emousl. ~known~SNP~ ~.~
c- ~~_ ~
_ . - x: 3 ,;x ..c r . ;. .~~.:. . - Wit... ,.;~, ; t .~ ' ~r w ~i_..H ~ ~ - .~~ s_ :_"'e' g'.n~ ~::;~,s~";f ,.~;. :W, ~ .:;;... C: ._.~ .a, -~r."~ .. P . ~-.a -.:., .:... R ~ ..,..~,~ -, m x--: _-~..~ .:~~"- ,x ~~ , ;5..~".<
use ~iienee =- su ~ ~ y~:v~ ~~~ _. ~ :-~~.~ y--., ~.'as.'"'.. w- ..,m'~.'..e:". .,~.~ . s~ ~ i,. ~,x ~"",..:. .,a~,s~",:a ~.,ir. r.,~~,~ ,~",; .. !i~ .. ~',~,..~.~1~.
~ 103 Q No 137 A -> No 84 N -> S Yes 1 -> No 203 R -> No 223 Y -> H Yes 273 V -> No 275 V -> No 285 K -> No 285 K -> Q No 311 E -> No 482 V -> A No Variant protein HSAPHOL P3 is encoded by the following transcript(s): HSAPHOL
T5, for which the sequences) is/are given at the end of the application. The coding portion of transcript HSAPHOL TS is shown in bold; this coding portion starts at position 253 and ends at position 1704. The transcript also has the following SNPs as listed in Table 8 (given according 10 to their position on the nucleotide sequence, with the alternative nucleic acid listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL P3 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 8 - Nucleic acid SNPs SNP position on nucleotideAlternative nucleic Previously.known acid SNP?
se uence. :~ ~ ~ ~,~ ~
q r ~~k -r ~.~ ~..~~:~- ~ : ~ a ,: ~
..._so - ..__. _ .. . ~ e_ 179 G -> C No 231 A -> No 1071 G -> No 1077 G -> No 1105 A -> No 1105 A -> C No 1185 G -> No 1554 C -> T Yes 1674 G -> T Yes 1697 T -> C No 1772 C -> T Yes 1833 G -> A Yes 232 A -> T No 1940 A -> C Yes 2095 C -> T Yes 2140 A -> G Yes 2285 G -> No 2392 -> A No 2409 T -> G No 281 T -> No 462 C -> T Yes 503 A -> G Yes 561 G -> No 662 C -> No 860 G -> No 919 T -> C Yes Variant protein HSAPHOL P4 according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) HSAPHOL_T6. An alignment is given to the known protein (Alkaline phosphatase, tissue-nonspecific isozyme precursor) at the end of the application. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between HSAPHOL P4 and AAH21289:
l.An isolated chimerie polypeptide encoding for HSAPHOL P4, comprising a first amino acid sequence being at least 90 % homologous to MGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLC
GVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAA
YAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDV
EYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEP
GDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALH
EAVEMDRAIGQAGSLTSSEDTLTWTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKP
FTAILYGNGPGYKV VGGERENV SMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGP
MAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVL
F corresponding to amino acids 124 - 586 of AAH21289, which also corresponds to amino acids 1 - 463 of HSAPHOL P4.
Comparison report between HSAPHOL P4 and PPBT HUMAN
l .An isolated chimeric polypeptide encoding for HSAPHOL P4, comprising a first amino acid sequence being at least 90 % homologous to MGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLC
GVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAA
YAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDV
EYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEP
GDMQYELNRNNVTDPSLSEMVWAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALH
EAVEMDRAIGQAGSLTSSEDTLTWTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKP
FTAILYGNGPGYKWGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGP
MAHLLHGVHEQNYVPH VMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVL
F corresponding to amino acids 62 - 524 of PPBT HUMAN, which also corresponds to amino acids 1 - 463 of HSAPHOL P4.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
membrane. The protein localization is believed to be membrane because only one of the two trans-membrane region prediction programs (Tmpred: l, Tmhmm: 0) has predicted that this protein has a trans-membrane region, but similarity to known proteins suggests a GPI anchor. In addition both signa~peptide prediction programs predict that this protein is a non-secreted protein..
Variant protein HSAPHOL P4 also has the following norrsilent SNPs (Single Nucleotide Polymorphisms) as listed in Table 9, (given according to their positions) on the amino acid 1 S sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL_P4 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 9 - Amino acid mutations t f.,., ~. ~C~~~' 7F4'n -.- ",.~t9 ~f :.,'" "~ , b ;.: a,~-...:. c. .~
~.,~ ~ ~a"~,....,.xm,". . .~..~,Eea$,.".. ~:a~ <ir~~".. i ~~
a~ s ~.x."." ~'.xu,~.~y";. ,-.~.
.. ~a .. c"=, ' ~ ~' . ~1~~'~m~tb~~.Cl~.-A,~ x:~a .~: a . re~~ously~ o o~ SlslF~
p , ( , ~ . ~~a ~ ~ t -;x x - ~W ~~~w ~..~,... ~.~tetatye4 a~n~no baGxd ;s~ .
-~~~r~ ~. a "' ~'. ,~ ~ ~,, ~ ~ as sequence= ~c ~
W ~ a ~,.. .~ '.'~~.... ~ ~'it-w . .:c~:~''~'~'"~~'~r~~~~x~~ x ~ ~s~, .,~ No ~
116 A ->
182 R -> No 82 Q -> No 202 Y -> H Yes 252 V -> No 254 V -> No 264 K -> No 264 K -> Q No 290 E -> No 461 V -> A No 63 J N -> S Yes Variant protein HSAPHOL P4 is encoded by the following transcript(s):
HSAPHOL_T6, for which the sequences) is/are given at the end of the application. The coding portion of transcript HSAPHOL-T6 is shown in bold; this coding portion starts at position 215 and ends at position 1603. The transcript also has the following SNPs as listed in Table 10 (given according to their position on the nucleotide sequence, with the alternative nucleic acid listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL P4 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 10 - Nucleic acid SNPs ,w r SNP ' ~h n~ ~. .~~, .,, ~.~ , . ~z, ... :~.~ ~ >
pos ~ o ~.o~ nueleQ.t~de . Alte -rnah~~-nu~'':~
:~~:~ ~,; ~ c exc. acrd: ee ;m zsl kn - wn -~ Y . o SN m .~
w -.~~. :m-~.~ ~ ~ =-. se ue c . , ~ o~ ~v ,.
Y 3:
~,~.
z ~..: ..a ~~ F ~ ::~.. ,x.s , ~;zs a ".,. "paYy"
~~,',.~r...m;.: . r... . ~ w".,: ' ~ .v;-.~'.. <.~'',~ ~:.~~=
361 C - j T Yes 402 A -> G Yes 1084 G _>
No 1453 C -> T
Yes 1573 G -> T
Yes 1596 T -> C No 1671 C -> T
Yes 1732 G -> A
Yes 1839 A -> C Yes 1994 C -> T Yes 2039 A -> G Yes 2184 G _>
No 460 G -> No 2291 -> A No 2308 T -> G No 561 C -> No 759 G -> No 818 T -> C Yes 970 G -> No 976 G -> No 1004 A -> No 1004 A -> C No Variant protein HSAPHOL PS according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) HSAPHOL_T7. An alignment is given to the known protein (Alkaline phosphatase, tissue-nonspecific isozyme precursor) at the end of the application. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between HSAPHOL PS and AAH21289:
l.An isolated chimeric polypeptide encoding for HSAPHOL P5, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
GLFEPGDMQYELNRNNVTDPSLSEMV V VAIQILRKNPKGFFLLVEGGRIDHGHHEGKA
KQALHEAVEM corresponding to amino acids 63 - 417 of AAH21289, which also corresponds to amino acids 1 - 355 of HSAPHOL PS, and a second amino acid sequence being at least 90 homologous to DHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVD
YAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIG
ANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 440 - 586 of AAH21289, which also corresponds to amino acids 356 - 502 of HSAPHOL P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated chimeric polypeptide encoding for an edge portion of I-ISAPHOL-P5, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise MD, having a structure as follows: a sequence starting from any of amino acid numbers 355-x to 355; and ending at any of amino acid numbers 356+ ((n-2) - x), in which x varies. from 0 to n-2.
Comparison report between HSAPHOL PS and PPBT HUMAN
l.An isolated chimeric polypeptide encoding for HSAPHOL P5, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGV STVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
GLFEPGDMQYELNRNNVTDPSLSEMVWAIQILRKNPKGFFLLVEGGRIDHGHHEGKA
KQALHEAVEM corresponding to amino acids 1 - 355 of PPBT HUMAN, which also corresponds to amino acids 1 - 355 of HSAPHOL P5, and a second amino acid sequence being at least 90 % homologous to DHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVD
YAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIG
ANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 377 - 524 of PPBT HUMAN, which also corresponds to amino acids 356 - 502 of HSAPHOL_P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated chimeric polypeptide encoding for an edge portion of HSAPHOL_P5, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise MD, having a structure as follows: a sequence starting from any of amino acid numbers 355-x to 355; and ending at any of amino acid numbers 356+ ((n-2) - x), in which x varies from 0 to rr2.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
membrane. The protein localization is believed to be membrane because of manual inspection of known protein localization and/or gene structure and/or similarity to known protein..
Variant protein HSAPHOL PS also has the following norrsilent SNPs (Single Nucleotide Polymorphisms) as listed in Table I 1, (given according to their positions) on the amino acid sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL PS
sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table ll -Amino acid mutations »- ., .~W x - ~~.W~
~S : ° ositso s, ~=on~amrno. aci .
P ~(.,~ ..,~ d~ Alternative arriino acrd ~ Prevro - -1:
~~~ r. ~ ~~ .; ~,~, , ~ a .uS y 1C110Wn S' ~4.~
.r ~ri ~..~'"~ A*."~
~ x~ ~ ° :~: ;~, ~: ~ m =~ :~: ~'.
124 ~ N -> S Yes 143 Q -> No 500 V -> A No 10 I -> No 1 ~~ A -> No 243 R -> No 263 Y -> H Yes 313 V -> No 315 V -> No 325 K -> No 325 K -> Q No 351 E -> No IS
DEMANDE OU BREVET VOLUMINEUX
LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND
PLUS D'UN TOME.
NOTE : Pour les tomes additionels, veuillez contacter le Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS
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NOTE: For additional volumes, please contact the Canadian Patent Office NOM DU FICHIER / FILE NAME
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Gland Gland of 24 64-Am-Kidney 111PO1O1Bmbion Kidney PM-Pool M8~F
of 14 65-CI-Kidney 1110970 ClontechKidney PM-Pool M&F
of 14 66-B-Kidney A411080 BiochainKidney PM-Pool M&F
of 5 67-CG-CerebellumCG-183-5IchilovCerebellum PM M/74 68-CG-CerebellumCG-212-5IchilovCerebellum PM M/54 69-B-Brain A411322 BiochainBrain PM M/28 70-CI-Brain 1120022 ClontechBrain PM-Pool M&F
of 2 71-B-Brain A411079 BiochainBrain PM-Pool M&F
of 2 72-CG-Brain CG-151-IIchilovBrain PM F/86 73-Am-Skeletal 1O1P013Ambion Skeletal PM F/28 Muscle Muscle 74-CI-Skeletal 1061038 ClontechSkeletal PM-Pool M&F
Muscle Muscle of 2 Materials and Experimental Procedures RNA preparation - RNA was obtained from Clontech (Franklin Lakes, NJ USA
07417, www.clontech.com), BioChain Inst. Inc. (Hayward, CA 94545 USA
www.biochain.com), ABS
(Wilmington, DE 19801, USA, http://www.absbioreagents.com) or Ambion (Austin;
USA, http://www.ambion.com). Alternatively, RNA was generated from tissue samples using TRI-Reagent (Molecular Research Center), according to Manufacturer's instructions. Tissue and RNA samples were obtained from patients or from postmortem. Total RNA samples were treated with DNaseI (Ambion) and purified using RNeasy columns (Qiagen).
RT PCR - Purified RNA (1 pg) was mixed with I50 ng Random Hexamer primers (Invitrogen) and 500 pM dNTP in a total volume of 15.6 pl. The mixture was incubated for 5 min at 65 °C and then quickly chilled on ice. Thereafter, 5 p.l of SX
SuperscriptII first strand buffer (Invitrogen), 2.4p,1 O.1M DTT and 40 units RNasin (Promega) were added, and the mixture was incubated for 10 min at 25 °C, followed by further incubation at 42 °C for 2 min.
Then, 1 p l (200units) of SuperscriptII (Invitrogen) was added and the reaction (final volume of 25p1) was incubated for 50 min at 42 °C and then inactivated at 70 °C for ISmin. The resulting cDNA was diluted 1:20 in TE buffer (10 mM Tris pH=8, 1 mM EDTA pH=8).
Real-Time RT-PCR analysis- cDNA (Spl), prepared as described above, was used as a template in Real-Time PCR reactions using the SYBR Green I assay (PE Applied Biosystem) with specific primers and UNG Enzyme (Eurogentech or ABI or Roche). The amplification was effected as follows: 50 "C for 2 min, 95 °C for 10 min, and then 40 cycles of 95 "C for l5sec, followed by 60 °C for 1 min. Detection was performed by using the PE
Applied Biosystem SDS
7000. The cycle in which the reactions achieved a threshold level (Ct) of fluorescence was registered and was used to calculate the relative transcript quantity in the RT reactions. The relative quantity was calculated using the equation Q=efficiency~-~'. The efficiency of the PCR
reaction was calculated from a standard curve, created by using serial dilutions of several reverse transcription (RT) reactions. To minimize inherent differences in the RT reaction, the resulting relative quantities were normalized to the geometric mean of the relative quantities of several housekeeping (HSKP) genes. Schematic summary of quantitative real-time PCR
analysis is presented in Figure 3. As shown, the x axis shows the cycle number. The CT =
Threshold Cycle point, which is the cycle that the amplification curve crosses the fluorescence threshold that was set in the experiment. This point is a calculated cycle number in which PCR
products signal is above the background level (passive dye ROX) and still in the Geometric/Exponential phase (as shown, once the level of fluorescence crosses the measurement threshold, it has a geometrically increasing phase, during which measurements are most accurate, followed by a linear phase and a plateau phase; for quantitative measurements, the latter two phases do not provide accurate measurements). The taxis shows the normalized reporter fluorescence. It should be noted that this type of analysis provides relative quantification.
The sequences of the housekeeping genes measured in all the examples on ovarian cancerpanel were as follows:
SDHA (GenBank Accession No.1VM-004168) SDHA Forward primer: TGGGAACAAGAGGGCATCTG
SDHA Reverse primer: CCACCACTGCATCAAATTCATG
SDHA-amplicon TGGGAACAAGAGGGCATCTGCTAAAGTTTCAGATTCCATTTCTGCTCAGTATCCAGT
AGTGGATCATGAATTTGATGCAGTGGTGG
PBGD (GenBank Accession No. BC019323), PBGD Forward primer: TGAGAGTGATTCGCGTGGG
PBGD Reverse primer: CCAGGGTACGAGGCTTTCAAT
PBGD-amplicon:
TGAGAGTGATTCGCGTGGGTACCCGCAAGAGCCAGCTTGCTCGCATACAGACGGAC
AGTGTGGTGGCAACATTGAAAGCCTCGTACCCTGG
HPRT1 (GenBank Accession No. NM 000194), HPRT1 Forward primer: TGACACTGGCAAAACAATGCA
HPRT1 Reverse primer: GGTCCTTTTCACCAGCAAGCT .
HPRT1-amplicon:
TGACACTGGCAAAACAATGCAGACTTTGCTTTCCTTGGTCAGGCAGTATAATCCAA
AGATGGTCAAGGTCGCAAGCTTGCTGGTGAAAAGGACC
GAPDH (GenBank Accession No. BC026907) GAPDH Forward primer: TGCACCACCAACTGCTTAGC
GAPDH Reverse primer: CCATCACGCCACAGTTTCC
GAPDH-amplicon TGCACCACCAACTGCTTAGCACCCCTGGCCAAGGTCATCCATGACAACTTTGGTATC
GTGGAAGGACTCATGACCACAGTCCATGCCATCACTGCCACCCAGAAGACTGTGGA
TGG
The sequences of the housekeeping genes measured in all the examples on normal tissue samples panel were as follows:
RPL19 (GenBank Accession No. NM_000981), RPL19 Forward primer: TGGCAAGAAGAAGGTCTGGTTAG
RPL19 Reverse primer: TGATCAGCCCATCTTTGATGAG
RPL19 -amplicon:
TGGCAAGAAGAAGGTCTGGTTAGACCCCAATGAGACCAATGAAATCGCCAATGCCA
ACTCCCGTCAGCAGATCCGGAAGCTCATCAAAGATGGGCTGATCA
TATA box (GenBank Accession No. NM 003194), TATA box Forward primer : CGGTTTGCTGCGGTAATCAT
TATA box Reverse primer: TTTCTTGCTGCCAGTCTGGAC
TATA box -amplicon:
CGGTTTGCTGCGGTAATCATGAGGATAAGAGAGCCACGAACCACGGCACTGATTTT
CAGTTCTGGGAAAATGGTGTGCACAGGAGCCAAGAGTGAAGAACAGTCCAGACTG
GCAGCAAGAAA
Ubiquitin (GenBank Accession No. BC000449) Ubiquitin Forward primer: ATTTGGGTCGCGGTTCTTG
Ubiquitin Reverse primer: TGCCTTGACATTCTCGATGGT
Ubiquitin C -amplicon:
ATTTGGGTCGCGGTTCTTGTTTGTGGATCGCTGTGATCGTCACTTGACAATGCAGAT
CTTCGTGAAGACTCTGACTGGTAAGACCATCACCCTCGAGG
TTGAGCCCAGTGACACCATCGAGAATGTCAAGGCA
SDHA (GenBank Accessio n No. NM 004168) SDHA Forward primer:
TGGGAACAAGAGGGCATCTG
SDHA Reverse primer: CCACCACTGCATCAAATTCATG
SDHA-amplicon TGGGAACAAGAGGGCATCTGCTAAAGTTTCAGATTCCATTTCTGCTCAGTATCCAGT
AGTGGATCATGAATTTGATGCAGTGGTGG
Oligonucleotide-based micro-array experiment protocol-Microarray fabrication Microarrays (chips) were printed by pin deposition using the MicroGrid II MGII
robot from BioRobotics Limited (Cambridge, UK). 50-mer oligonucleotides target sequences were designed by Compugen Ltd (Tel-Aviv, IL) as described by A. Shoshan et al, "Optical technologies and informatics", Proceedings of SPIE. Vol 4266, pp. 86-95 (2001). The designed oligonucleotides were synthesized and purified by desalting with the Sigma-Genosys system (The Woodlands, TX, US) and all of the oligonucleotides were joined to a C6 amino-modified linker at the 5' end, or being attached directly to CodeLink slides (Cat #25-6700-O1. Amersham Bioscience, Piscataway, NJ, US). The 50-mer oligonucleotides, funning the target sequences, were first suspended in Ultra-pure DDW (Cat # O1-866-lA Kibbutz Beit-Haemek, Israel) to a concentration of SOpM. Before printing the slides, the oligonucleotides were resuspended in 300mM sodium phosphate (pH 8.5) to final concentration of 150mM and printed at 35-40%
relative humidity at 21 °C.
Each slide contained a total of 9792 features in 32 subarrays. Of these features, 4224 features were sequences of interest according to the present invention and negative controls that were printed in duplicate. An additional 288 features (96 target sequences printed in triplicate) contained housekeeping genes from Human Evaluation Library2, Compugen Ltd, Israel.
Another 384 features are E.coli spikes 1-6, which are oligos to E Coli genes which are commercially available in the Array Control product (Array control- sense oligo spots, Ambion Inc. Austin, TX. Cat #1781, Lot #112K06).
Post-coupling processing of printed slides After the spotting of the oligonucleotides to the glass (CodeLink) slides, the slides were incubated for 24 hours in a sealed saturated NaCI humidification chamber (relative humidity 70-75%).
Slides were treated for blocking of the residual reactive groups by incubating them in blocking solution at 50°C for 15 minutes (lOml/slide of buffer containing O.1M Tris, SOmM
ethanolamine, 0.1% SDS). The slides were then rinsed twice with Ultra-pure DDW
(double distilled water). The slides were then washed with wash solution (lOml/slide.
4X SSC, 0.1%
SDS)) at 50°C for 30 minutes on the shaker. The slides were then rinsed twice with Ultra-pure DDW, followed by drying by centrifugation for 3 minutes at 800 rpm.
Next, in order to assist in automatic operation of the hybridization protocol, the slides were treated with Ventana Discovery hybridization station barcode adhesives.
The printed slides were loaded on a Bio-Optica (Milan, Italy) hematology staining device and were incubated for 10 minutes in SOmI of 3-Aminopropyl Triethoxysilane (Sigma A3648 lot #122K589). Excess fluid was dried and slides were then incubated for three hours in 20 mm/Hg in a dark vacuum desiccator (Pelco 2251, Ted Pella, Inc. Redding CA).
The following protocol was then followed with the Genisphere 900-RP (random primer), with mini elute columns on the Ventana Discovery HybStationT"'', to perform the microarray experiments. Briefly, the protocol was performed as described with regard to the instructions and information provided with the device itself. The protocol included cDNA
synthesis and labeling. cDNA concentration was measured with the TBS-380 (Turner Biosystems.
Sunnyvale, CA.) PicoFlour, which is used with the OliGreen ssDNA Quantitation reagent and kit.
Hybridization was performed with the Ventana Hybridization device, according to the provided protocols (Discovery Hybridization Station Tuscon AZ).
The slides were then scanned with GenePix 4000B dual laser scanner from Axon Instruments Inc, and analyzed by GenePix Pro 5.0 software.
Schematic summary of the oligonucleotide based microarray fabrication and the experimental flow is presented in Figures 4 and 5.
Briefly, as shown in Figure 4, DNA oligonucleotides at 25uM were deposited (printed) onto Amersham 'CodeLink' glass slides generating a well defined 'spot'. These slides are covered with a long-chain, hydrophilic polymer chemistry that creates an active 3-D surface that covalently binds the DNA oligonucleotides 5'-end via the C6-amine modification. This binding ensures that the full length of the DNA
oligonucleotides is available for hybridization to the cDNA and also allows lower background, high sensitivity and reproducibility.
Figure 5 shows a schematic method for performing the microarray experiments.
It should be noted that stages on the le$-hand or right-hand side may optionally be performed in any order, including in parallel, until stage 4 (hybridization). Briefly, on the left-hand side, the target oligonucleotides are being spotted on a glass microscope slide (although optionally other materials could be used) to form a spotted slide (stage 1). On the right hand side, control sample RNA and cancer sample RNA are Cy3 and Cy5 labeled, respectively (stage 2), to form labeled probes. It should be noted that the control and cancer samples come from corresponding tissues (for example, normal prostate tissue and cancerous prostate tissue).
Furthermore, the tissue from which the RNA was taken is indicated below in the specific examples of data for particular clusters, with regard to overexpression of an oligonucleotide from a "chip"
(microan-ay), as for example "prostate" for chips in which prostate cancerous tissue and normal tissue were tested as described above. In stage 3, the probes are mixed. In stage 4, hybridization is performed to form a processed slide. In stage 5, the slide is washed and scanned to form an image file, followed by data analysis in stage 6.
Cluster H61775 features 2 transcripts) and 6 segments) of interest, the names for which are given in Tables 1 and 2, respectively, the sequences themselves are given at the end of the application. The selected protein variants are given in table 3.
Table I - Transcripts of interest .Transcn .~ .;~-tName: .:~ 0~
'~ ~~ . ,~ SEQ-.~I
. ~ ,, D N
~p ~ ~--=.
..
-~~ ~:~~
=
H61775 T21 _ . ,, , . ..:
... .
Table 2 - Segments of interest --Segment ame ~-~~~ ~-SE ~ - ~ ~ T~:;~ w.~.~~
- ~ ~ ~ ~ ~ ~~r Q I D NO..
H61775 node 2 3 .:
~w ~ ~ ..~...~ .
~
H61775 node 4 H61775 node 6 H61775 node-8 6 H61775 node 0 7 H61775 node 5 g Table 3 - Proteins of interest otein -.
~ ~~ ~ ~ ~~
ame ~
~
. .
z ,.
~ , . ~,. , ;t, ' .'ts. ,...~ ~ ~n ., ix ~~ ~a. , ~'~ - ~, ~ ,~:'f~ z.~ ~~!1~~:.,.
~., ~~ H~
. a ,.,., .~x, 5 ~6 4 7 ~.;
~uh~..m~ , ~.a1 i..~ ~
~ :
~
H61775 P16 .,w .
..
~~ ~Lz. - N..~'.
Cluster H61775 can be used as a diagnostic marker according to overexpression of transcripts of this cluster in cancer. Expression of such transcripts in normal tissues is also given according to the previously described methods. The term "number" in the right hand column of the table and the numbers on the taxis of Figure 6 refer to weighted expression of ESTs in each category, as "parts per million" (ratio of the expression of ESTs for a particular cluster to the expression of all ESTs in that category, according to parts per million).
Overall, the following results were obtained as shown with regard to the histograms in Figure 6 and Table 4. This cluster is overexpressed (at least at a minimum level) in the following pathological conditions: brain malignant tumors and a mixture of malignant tumors from different tissues.
Table 4 - Normal tissue distribution Nam' o fTzssue _Number bladder 0 brain 0 colon 0 epithelial 10 general 3 breast g muscle 0 ovary 0 pancreas 0 prostate 0 uterus 0 Table S - P values and rat ios for expression in cancerous tissue r ~ u~. ~ ~~ P
r Name: of ~ ' ''= ~y -'~.~~SP2 R;4 Tissue. p f 2_~~x '~~ R3 .- ' ' SP1: ~ fi _ ek~.'I,~
~ % ' .._ . ., ~ ;. Y" ~ .",.
~rv ,.
~bladder~m ~3 E~.,S~c"..'firs . - , ..
R~
~O
. 3.8e-O13.2e-O12.5 4.6e-O1 1.9 e-l brain 8.8e-026.Se-021 3.5 4.1e-04 5.8 colon 5.6e-O16.4e-O11 1.1 1 1.1 epithelial 3.Oe-021.3e-O12.3e-022.1 3.2e-O1 1.2 general 1.3e-064.9e-OSl.Oe-076.3 l.Se-06 4.3 breast 4.7e-01 3.7e-O13.3e-O12.0 4.6e-O11.6 muscle 2.3e-O1 2.9e-O11.5e-O16.8 3.9e-O12.6 ovary 3.8e-O1 4.2e-O11.5e-012.4 2.6e-Ol1.9 pancreas 3.3e-O1 4.4e-Ol4.2e-O12.4 5.3e-O11.9 prostate 7.3e-O1 7.8e-OI6.7e-OI1.5 7.Se-O11.3 uterus 1.Oe-01 2.6e-O12.9e-O 2.6 5. I 1.8 l e-O1 As noted above, cluster H61775 Ieatures 2 transcript(s), which were listed in Table 1 above. A description of each variant protein according to the present invention is now provided.
Variant protein H61775 P16 according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) H61775 T21. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between H61775 P16 and Q9P2J2 (SEQ ID N0:953):
l.An isolated chimeric polypeptide encoding for H61775 P16, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 11 - 93 of Q9P2J2, which also corresponds to amino acids 1 - 83 of H61775 P16, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV corresponding to amino acids 84 - 152 of H61775 P16, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated polypeptide encoding for a tail of H61?75 P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV in H617?5 P16.
Comparison report between H61775 P16 and AAQ88495 (SEQ ID N0:954):
I .An isolated chimeric polypeptide encoding for I-161775 P 16, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 1 - 83 of AAQ88495, which also corresponds to amino acids 1 - 83 of H61775 P16, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the~sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV corresponding to amino acids 84 - 152 of H61775 P 16, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated polypeptide encoding for a tail of H61775 P16, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95% homologous to the sequence DCGFPAFRELKRAETVSPVFFTRRCIWEDLKSTGFSPAGGGRPPGGGPRTQEDSGLPCW
RSSCSVTLQV in H61775 P16.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
secreted. The protein localization is believed to be secreted because both signal-peptide prediction programs predict that this protein has a signal peptide, and neither trans-membrane region prediction program predicts that this protein has a trans-membrane region..
Variant protein H61775 P16 also has the following norrsilent SNPs (Single Nucleotide Polymorphisms) as listed in Table 6, (given according to their positions) on the amino acid sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein H61775 P 16 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 6 - Amino acid mutations :. .,>cf ~~~.~'4~~w;?~~K~, n ~. Alternatue~ain~no~acid~ Pieviousl ;=known SNP 'ositiori s -Son:ayuno~acids 'i SNP?
p ~ ) ~ b - ( ) Y
;~. . , ., --~,:..E ~ ~ ~ ~ ~
. , ~r~ : <
~
~
~se uence ~: ~. , ~ .~~
, ~~- .: ~ ~ . v;
_ .. ~
zq P. ,~ -~~ ~ ~ . ~~ ., : . .~ ..
~ ~ . ~ ~ ~ z , ~"~.
~. t-x~ .~ _..~.~ ~_. . ,_ ~~ ___~ N~ , ~ .w .. ...__ 14 I -> T No 138 G -> R No 34 G -> E Yes 48 G -> R No 91 R -> * Yes Variant protein H61775 P16 is encoded by the following transcript(s): H61775 T21, for which the sequences) is/are given at the end of the application. The coding portion of transcript H61775 T21 is shown in bold; this coding portion starts at position 261 and ends at position 716. The transcript also has the following SNPs as listed in Table 7 (given according to their position on the nucleotide sequence, with the alternative nucleic acid listed;
the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein H61775 P16 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 7 - Nucleic acid SNPs a., :PfY' satFwv'.,.~.. . ":..~:rn .yd~, . ..f.:. '~, .iff0.,yx., "~ ~,,x ~..-. y : _ a , . : ,m ~i '.
z. nucleotide ~'. ~;Altemat ~ uel . e- acidp - ~ .ev ou 1~ .known. SNP
S ~,pas~han ova .. ~ e~..; ~n ~ ' o y , TTws -._ ' ' ~ ~,, ~F,,.,.w .' sx'."' :e:~.cai. z ~.,re ,. x,. 'fi" '~ arY, ,i m,..,.r ~~,"sm,~
equ~nc~ .. ~ ~ ~. ate. - ~ ~ r- .: ~s " ,. ~.
" ,"i~.",~, .." w.='~.. ,..,f'. x.r~' ..~~,4:::..a.. .~.r~,_ : ~R~. ~,: m~<., ~ ~~cr;, °a . .
117 T -> C Yes 200 T -> C No 672 G -> C No 222 T -> C Yes 301 T -> C No 361 G -> A Yes 377 G -> A No 400 -> C No 402 G -> C No 531 C -> T Yes 566 ~ T -> C ~ No Variant protein I-161775 P17 according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) H61775 T22. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between H61775 P17 and Q9P2J2:
l.An isolated chimeric polypeptide encoding for H61775 P17, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 11 - 93 of Q9P2J2, which also corresponds to amino acids 1 - 83 of H61775 P17.
Comparison report between H61775 P17 and AAQ88495:
l.An isolated chimeric polypeptide encoding for H61775 P17, comprising a first amino acid sequence being at least 90 % homologous to MVWCLGLAVLSLVISQGADGRGKPEVVSVVGRAGESVVLGCDLLPPAGRPPLHVIEWL
RFGFLLPIFIQFGLYSPRIDPDYVG corresponding to amino acids 1 - 83 of AAQ88495, which also corresponds to amino acids 1 - 83 of H61775 P17.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
secreted. The protein localization is believed to be secreted because both signal-peptide prediction programs predict that this protein has a signal peptide, and neither trans-membrane region prediction program predicts that this protein has a trans-membrane region..
Variant protein H61775 P17 also has the following norrsilent SNPs (Single Nucleotide Polymorphisms) as listed in Table 8, (given according to their positions) on the amino acid sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein H61775 P17 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 8 - Amino acid mutations 3 7k"' '::' RE- -r ...c '.,>: _ r ~.".- - '~'...
'.' ~~~~?. -~. . , 'n _,.. T~:~e"~"
.,..2 3v'..~ri c~ ,k9~ f" ~"w .' .. vy~-35".
7 ° '.:. ~5 ..,..h~ ., .3.,. ,:.~ ,..~a' , , ~,~, .~.t,' ..;t" n ,.l.y- ~.".o. .; .~;, M,..,.,. .. 2 . Y~e.~Fy: -~
~4 ' , ' ,: % ' "..i,'", ... 'GSS -.y . x. dl v:~F°a~~_..
osati n .s ~on~ammo~acid F..Alternatiue~ammo.,acid s= m I'xemousl ~'known,-SNP
. x".
,k..h'.'a~w, ". "'& ~ 'f'°.. r w~'~:3F
R ". ~ -:.. ,."~.Xt' .. ~....3~°' ~~ ~.:~~ ~ ~z '-2~Y~W~~ -.:,W~ -. 'SYZ" W: ~. n~..-- __.",w I -.;~ rte.
'~RL ' ">~~~~ ', ~ ~ , , ~,.,'3 f.: .~ m~ s,,w Y'". Fi : ~, % aFSf ."-..,~, .. , t~~~ , V ~ -, s L.. S::":, " ~ . ., 5 ~s'' ,~Y . , S.f. ' ,,~y"~'T-.~,.
'K
_~;,x2 I .:.>~s:.:", ,:b~
-~se uencer. ~,~T_,. "
,..,y. ' -~'. .,'-.. S~m,. , ,z sr,~ ~x...~,~..,..:'~... terse. ~a', ,.._r ~.-~-:~- .~.~ ~, Wig...-.~,,~.: v- '~. ,;~::~y:~;.- ~,-<..
..,d~" ~..., " ~ ' "~Y,...,.,.3"t'.r N .,~~ ,... ,. x'=~,>w.~~,.;.,"
14 "xI ->" T " No 34 G -> E Yes 48 G -> R No Variant protein H61775 P17 is encoded by the following transcript(s): H61775 T22, for which the sequences) is/are given at the end of the application. The coding portion of transcript H61775 T22 is shown in bold; this coding portion starts at position 261 and ends at position 509. The transcript also has the following SNPs as listed in Table 9 (given according to their position on the nucleotide sequence, with the alternative nucleic acid listed;
the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein H61775 P17 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 9 - Nucleic acid SNPs ~ :w,~ ~ ~ - ~ ~ r ~_~~"F,.,.< . ... :° ~:; !~
~.'F
r "1 ~ id .;, :eYi = : 1 ,s NPR ~ osiiwton on.nu~l"eatrde , _ Iteriiatxve~nuc a c c ~ 9 x out. . _I~no~ S
,b ~ . a =.sequence F ~ ~ ,~ a.~ w ~~ .
.,_ _ .,,>W ~-.,.. ~~ r.,. ~ w;
117 T -> C ~ Yes 200 T -> C No 222 T -> C Yes 301 T -> C No 361 G -> A Yes 377 G -> A No 400 -> C No 402 G -> C No 596 T -> A Yes Ac nc,ted above. cluster61775 features fi H se~ment(sl. which were listed in Table 2 above and for which the sequences) are given at the end of the application. These segments) are portions of nucleic acid sequences) which are described herein separately because they are of particular interest. A description of each segment according to the present invention is now provided.
Segment cluster H61775 node 2 according to the present invention is supported by 17 libraries. The number of libraries was determined as previously described.
This segment can be found in the following transcript(s): H61775 T21 and H61775 T22. Table 10 below describes the starting and ending position of this segment on each transcript.
Table 10 - Segment location on transcripts 'S ~~~;,.at ~a~~ ~"'~~'.. _ ~*. a : y = s'.
,nyaPH..,.,.,.u. .. ~t~ -.. .~, ..~y , 5'~~, ~~ y.. ~Se . n end~n asz~on s a . ent~ last n . g : tt, a ~g ~Traaascn ,.name ~' . osxlran- .~ ~~iro, ~ ,5 gm.~ ~. : ~....~_~a'.
~ ~.._.,. ~
;a~M ~., rya =5 ~~'"~;ar ' ~ - ~
bY '-- ~
~ '-sF s , .. ..
.~ .. .. .....~ .
m , , ,. ..: ,4u;a. .
",rz~. ...~,.,~s... ~ c .
.._ , ,fir ...- "a.3:.: ~, ~:dr ~' -~.""a. .
s .~:'~ ;.. r- S~e. ';- ,?~
-~' ~ , . ":.s~.
s ~-. ,~~,.' "~ .o.
~ ~" e ' ~'z~~ - x : -:~;'~
s~ ~
, ; : ,.
, . .,. .
a ,. .
. .. .
, .. x_ ...
. ..
Segment cluster H61775 node 4 according to the present invention is supported by 20 libraries. The number of libraries was determined as previously described.
This segment can be found in the following transcript(s): H61775 T21 and H61775 T22. Table 11 below describes the starting and ending position of this segment on each transcript.
Table 11 - Segment location on transcripts a,,. , ~ ~:~~ ~w v '~", ~Transe 't~riame;~ Se yen ~star=tin Se enl endln ~ ' osytton ,-~~ , . Lyon . h ~;t . _. ~ -~'I?~
~?p ~ ~ :~
~B P: ~ p _a -~
~ ~ ~~~ r ,,._~~
y ~-. _ _ ~. ~ : m_ .
>_ ~ ~, .. ~;,.
: ~ _~: ~ f. : ~ry - ~ ~ s ~ ' ~o :
~:~_: ~ ~~ ~s Segment cluster H61775 node 6 according to the present invention is supported by 1 libraries. The number of libraries was determined as previously described.
This segment can be found in the following transcript(s): H61775 T22. Table l2 below describes the starting and ending position of this segment on each transcript.
Table 12 - Segment location on transcripts ,~~ ~. ~ ~ ~ -X R J w .F. ~&. 'w~ ~I. , Transcn t<~iiame -~ ~~ " ent startnri S.e mrntwendm ~ .osvtson M. ositaon .W a..~
~ g p ~? . g g~l? ~ 5 ~
, , ., s ~ ~. ~
. .
~~
~
Segment cluster H61775 node 8 according to the present invention is supported by 5 libraries. The number of libraries was determined as previously described.
This segment can be found in the following transcript(s): H61775 T21. Table 13 below describes the starting and ending position of this segment on each transcript.
Table 13 - Segment location on transcripts rte: ~~- .. ~ .~ -~~;
~ -:~ - ~, .e. t ,. .x , ltl _ ~ _ ti ~a_ z ~p ~ n ..~. ~ - a _nt~a~dan ~ os~
- x. . ~e ent s ~z~t~ _ n ~~~~
Tra.~s ~ t,.name;: os a . g I?
. b= ;~~ v: ,. .
~ - :.
~ ~
~ na ~
H a , ~
~ ~ , ..~, .w.
~..~ . ~,: ~ ~
-~ ' .. ~ ~ ~ ~ f~ , "~ ' ~ .,r:
n~ ~
~~.:: ~;
. ~~. s _' ' ' -to :r.., S _b ~ , ..zm".~,.,.,. ~z "~ ~~
~ , - u__~.~.:
- Y ,re'.%.
According to an optional embodiment of the present invention, short segments related to the above cluster are also provided. These segments are up to about 120 by in length, and so are included in a separate description.
Segment cluster H61775 node 0 according to the present invention is supported by 4 libraries. The number of libraries was determined as previously described.
This segment can be found in the following transcript(s): H61775 T21 and H61775 T22. Table 14 below describes the starting and ending position of this segment on each transcript.
Table 14 - Segment location on transcripts H61775T2l I 86 Segment cluster H61775 node 5 according to the present invention can be found in the following transcript(s): H61775 T22. Table 15 below describes the starting and ending position of this segment on each transcript.
Table I S - Segment location on transcripts ~..1'.z2., ~2 .,rn.2,~- .~.d; ~. ~ .-~3r3.'~:-.wL:~-..~....:
'~-. ~ ~ . ,~ .....-3~,~" w"~,.
'T~. ,.~..>.~Yx ... - ~ '.~- i~F'x .. "..R
4.~'vr '~f" , ta~' Se' . entstartm ~ ..~Se ent endxn o~xtlon Transcn . t:~name~: osxtion,~; _, .:; ..y . ~~ , ~ .g P ~_ ~ p ~,~ ~? ~
~ ' ~
~__ .~. _. .7 a .,.~ -Fns ; :.~ ~"~~ ~, ~~ ..v~
' ,." ,.
.'.. ~ ~.--,~ .~, ~
-.n..~ ~_,. ~ - ~
~. : w -W
~~ ~a 6.~, ;~:.e5.
~_~. ty'-.- ~-:.,.,'.
:~'..,~~~5'" , ..,.. ..: w~%T ',~.CS
. L'.~'.~ ..,.s Variant protein alignment to the previously known protein:
Sequence name: /tmp/PswORJLCti/aLAXQjXh07:Q9P2J2 Sequence documentation:
Alignment of: H61775-P16 x Q9P2J2 ..
Alignment segment 1/1:
Quality: 803.00 Escore: 0 Matching length: 83 Total length: 83 Matching Percent Similarity: 100.00 Matching Percent Identity: 100.00 Total Percent Similarity: 100.00 Total Percent Identity: 100.00 Gaps: 0 Alignment:
IIIIIIIIIIIIIIIIillllllllllllllll 20 Sequence name: /tmp/PswORJLCti/aLAXQjXh07:AAQ88495 Sequence documentation:
Alignment of: H61775 P16 x AAQ88495 ..
Alignment segment 1/1:
Quality: 803.00 Escore: 0 Matching length: 83 Total length: 83 Matching Percent Similarity: 100.00 Matching Percent Identity: 100.00 Total Percent Similarity: 100.00 Total Percent Identity: 100.00 S Gaps: 0 Alignment:
IIIIiillllllllllllllllllllllllllllllllllllllilllll IIIIIIIIIIIIIIIIIIIIIIIiillllllll Sequence name: /tmp/naab8yR3GC/pSM412IL5o:Q9P2J2 Sequence documentation:
Alignment of: H61775 P17 x Q9P2J2 , .
Alignment segment 1/1:
Quality: 803.00 Escore: 0 Matching length: 83 Total length: 83 Matching Percent Similarity: 100.00 Matching Percent Identity: 100.00 Total Percent Similarity: 100.00 . Total Percent Identity: 100.00 Gaps: 0 Alignment:
IIIIIIIIIIIIIIIIilllllllllillllllilllllllillllllll IIIIIIIIIIIIIIIIIIilllllillllllli Sequence name: /tmp/naab8yR3GC/pSM412IL5o:AAQ88495 Sequence documentation:
Alignment of: H61775_P17 x AAQ88495 ..
Alignment segment 1/1:
Quality: 803.00 Escore: 0 Matching length: 83 Total length: 83 Matching Percent Similarity: 100.00 Matching Percent Identity: 100.00 Total Percent Similarity: 100.00 Total Percent Identity: 100.00 Gaps: 0 Alignment:
IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIII
IIIIIIIIIIIIIIIIIIIIIIillilllllll 25 Expression of immunoglobulin superfamily, member 9 H61775 transcripts which are detectable by amplicon as depicted in sequence name H61775seg8 in normal and cancerous ovary tissues.
Expression of immunoglobulin superfamily, member 9 transcripts detectable by or according to H61775seg8, H61775seg8 amplicon(s) and H61775seg8F2 and H61775seg8R2 primers was measured by real time PCR. In parallel the expression of four housekeeping genes:
PBGD (GenBank Accession No. BC019323; amplicon - PBGD-amplicon), HPRT1 (GenBank Accession No. NM 000194; amplicon - I-IPRT1-amplicon), and SDHA (GenBank Accession No. NM 004168; amplicon - SDHA-amplicon), GAPDI-I (GenBank Accession No.
BC026907;
GAPDI-1 amplicon) was measured similarly. For each RT sample, the expression of the above amplicon was normalized to the geometric mean of the quantities of the housekeeping genes.
The normalized quantity of each RT sample was then divided by the median of the quantities of the normal post-mortem (PM) samples (Sample Nos. 45-48,71, Table l, "Tissue samples in testing panel"), to obtain a value of fold up-regulation for each sample relative to median of the normal PM samples.
Figure 7 is a histogram showing over expression of the above-indicated immunoglobulin superfamily, member 9 transcripts in cancerous ovary samples relative to the normal samples.
(Values represent the average of duplicate experiments. Error bars indicate the minimal and maximal values obtained As is evident from Figure 7, the expression of immunoglobulin superfamily, member 9 transcripts detectable by the above amplicon(s) in cancer samples was significantly higher than in the norrcancerous samples (Sample Nos. 45-48, ,71 Table 1, "Tissue samples in testing panel") and including benign samples (samples No. 56, 62, 64). Notably an over-expression of at least 5 fold was found in 21 out of 43 adenocarcinoma samples.
Statistical analysis was applied to verify the significance of these results, as described below.
The P value for the difference in the expression levels of immunoglobulin superfamily, member 9 transcripts detectable by the above amplicon(s) in ovary cancer samples versus the normal tissue samples was determined by T test as 2.76E-4.
The above value demonstrates statistical significance of the results.
Primer pairs are also optionally and preferably encompassed within the present invention; for example, for the above experiment, the following primer pair was used as a norr limiting illustrative example only of a suitable primer pair: H61775seg8F2 forward primer; and H61775seg8R2 reverse primer.
The present invention also preferably encompasses any amplicon obtained through the use of any suitable primer pair; for example, for the above experiment, the following amplicon was obtained as a norrlimiting illustrative example only of a suitable amplicon: H61775seg8 H61775seg8F2 (SEQ ID N0:955) GAAGGCTCTTGTCACTTACTAGCCAT
H61775seg8R2 (SEQ ID N0:956) TGTCACCATATTTAATCCTCCCAA
Amplicon (SEQ ID N0:957) GAAGGCTCTTGTCACTTACTAGCCATGTGATTTTGGAAAGAAACTTAACATTAATTC
CTTCAGCTACAATGGAATTCTTGGGAGGATTAAATATGGTGACA
Expression of immunoglobulin superfamily, member 9 H61775 transcripts which are detectable by amplicon as depicted in sequence name H61775seg8 in different normal tissues.
Expression of immunoglobulin superfamily, member 9 transcripts detectable by or according to H61775 seg8 amplicon(s) and H61775 segBF and H61775 segBR was measured by real time PCR. In parallel the expression of four housekeeping genes -RPL19 (GenBank Accession No. NM 000981; RPL19 amplicon), TATA box (GenBank Accession No.
NM-003194; TATA amplicon), Ubiquitin (GenBank Accession No. BC000449; amplicon -Ubiquitin-amplicon) and SDHA (GenBank Accession No. NM 004168; amplicon - SDHA-amplicon) was measured similarly. For each RT sample, the expression of the above amplicon was normalized to the geometric mean of the quantities of the housekeeping genes. The normalized quantity of each RT sample was then divided by the median of the quantities of the ovary samples (Sample Nos. 18-20, Table 2 "Tissue samples in normal panel", above), to obtain a value of relative expression of each sample relative to median of the ovary samples.
The results are described in Figure 8, presenting the histogram showing the expression of H61775 transcripts which are detectable by amplicon as depicted in sequence name H61775seg8, in different normal tissues. Amplicon and primers are as above.
DESCRIPTION FOR CLUSTER HSAPHOL
Cluster HSAPHOL features 7 transcripts) and 18 segments) of interest, the names for which are given in Tables 1 and 2, respectively, the sequences themselves are given at the end of the application. The selected protein variants are given in table 3.
Table I - Transcripts of interest _~ ~ ~w, fw~~~ ~Cw.... ~ ~~, ~.'~rW.~,~_ ~"~.'v, T~ranscn txN me'~ ~ '~ .~ ...:',2'n' '&~"" p4~~'%..
_' ' ~--W ~SE ~ ~ID'~ O~' a ~ ,, ~~
s- - ~~. - ~ _ _ ~
~ ~ s s~'r ~.s"~' ~
T ra,~ ..a ~ ,."~~,a" " f~ '~~. .
~"~ ~. ,~$;r~ _ ~ . ~Y'r'9~~~'s,s~, , n a . , .-a k ~
Table 2 - Segments of interest .y-.,-. :z- .~; ~~ _ .' .C.. ~ -,~~ Rk ,.-a Y
Se~ a t'~T e~~. ~ ~:~' ~; ASE ~ ~II3x~NO~~ ~, ~ .>.
~ & ~ ~:~ ~x '.,~ x . w "'"<. kF . .~ 3 . ~b~ G~ : 7. "~x ~. i'r 7 _, ,' ~:1. 4,or i , ~:.'. .- ; .. ' 'u' ~ a. s ,~ , -4 ~ y ~~s.
~ ." ,A~~,.~ui- '4 HSAPHOL node 11 18 HSAPHOL node 13 19 HSAPHOL node 15 20 HSAPI-IOL node 19 21 HSAPI-IOL node 2 22 HSAPHOL node 21 23 HSAPI-IOL node 23 24 HSAPHOL node 26 25 HSAPI-IOL node 28 26 HSAPHOL node 38 27 HSAPHOL node 40 28 HSAPHOL node 42 29 HSAPHOL node 16 30 HSAPHOL node 25 31 HSAPHOL node 34 32 HSAPHOL node 35 33 HSAPHOL node 36 34 HSAPHOL node 41 ~ S
Table 3 - Proteins of interest ~roteua a~. ~ n~~Q ~D~~t~ .~, ~ ~~, ~,~
~r~ ~~ ~ a . ~m ''r~x s~ ~.+: a ~ -A
These sequences are variants of the known protein Alkaline phosphatase, tissue-nonspecific isozyme precursor (SwissProt accession identifier PPBT,HUMAN;
known also according to the synonyms EC 3.1.3.1; AP-TNAP; Liver/bone/kidney isozyme;
TNSALP), SEQ
ID NO: 36, referred to herein as the previously known protein.
The variant proteins according to the present invention are variants) of a known diagnostic marker, called Alkaline Phosphatase.
Protein Alkaline phosphatase, tissue-nonspecific isozyme precursor is known or believed to have the following funetion(s): THIS ISOZYME MAY PLAY A ROLE IN SKELETAL
MINERALIZATION. The sequence for protein Alkaline phosphatase, tissue-nonspecific isozyme precursor is given at the end of the application, as "Alkaline phosphatase, tissue-nonspecific isozyme precursor amino acid sequence". Known polymorphisms for this sequence are as shown in Table 4.
Table 4 - Amino acid mutations for Known Protein , SNPp,~osation(s)rori Comment ~».:~~ ~' .,,~~ ~, ima acid a nce . ~~~ ~ ~~ ~;~.
_.~_,t-1-. ~ >r. .~.~.;_:..,. ..~: -~.."~".....~',"w >s, ~..,w. ~;~ ,.
...~:.',-~, . : , ,,: .~a> ~u_ a.a ':~.w n".~....em~.,. '.. Via'':.'~.
28 Y -> C (in hypophosphatasia; infantile; 7% of activity).
/FTId=VAR 013972.
33 A -> V (in hypophosphatasia). /FTId=VAR 006147.
111 A -> T (in hypophosphatasia; odonto).
/FTId=VAR 006151.
116 A -> T (in hypophosphatasia; loss of activity).
/FTId=VAR 013977.
120 G -> R (in hypophosphatasia). /FTId=VAR 013978.
129 G -> R (in hypophosphatasia). /FTId=VAR 013979.
132 A -> V (in hypophosphatasia). /FTId=VAR 013146.
134 T -> N (in hypophosphatasia; 9% of activity).
/FTId=VAR 011082.
136 R -> H (in hypophosphatasia; moderate; 33% of activity).
/FTId=VAR 006152.
152 R -> H (in hypophosphatasia). /FTId=VAR O 13980.
162 G -> V (in hypophosphatasia; severe; 1 % of activity).
/FTId=VAR 006153.
170 N -> D (in hypophosphatasia). /FTId=VAR
013981.
40 A -> V (in hypophosphatasia; 2% of activity).
/FTId=VAR 011081.
171 H -> Y (in hypophosphatasia; severe;
2% of activity).
/FTId=VAR 006154.
I 76 A -> T (in hypophosphatasia). /FTId=VAR
0 t 1083.
177 A -> T (in hypophosphatasia; adult type).
/FTId=VAR 006155.
179 A -> T (in hypophosphatasia). /FTId=VAR
006156.
181 S -> L (in hypophosphatasia; 1 % OF
activity).
lFTId=VAR 013982.
184 R -> W (in hypophosphatasia; loss of activity).
/FTId=VAR 013983.
191 E -> G (in hypophosphatasia; odonto).
/FTId=VAR 006157.
191 E -> K (in hypophosphatasia; moderate;
frequent mutation in European countries). /FTId=VAR 006158.
201 C -> Y (in hypophosphatasia). /FTId=VAR
006159.
207 Q -> P (in hypophosphatasia). /FTId=VAR
006160.
51 A -> V (in hypophosphatasia). /FTId=VAR
013973.
211 N -> D (in hypophosphatasia). /FTId=VAR
013984.
220 G -> V (in hypophosphatasia; odonto).
/FTId=VAR 013985.
223 R -> W (in hypophosphatasia; 3% of activity).
/FTId=VAR 013986.
224 K -> E (in hypophosphatasia; infantile;
partial loss of activity). /FTId=VAR 011084.
235 E -> G (in hypophosphatasia). /FTId=VAR
013987.
246 R -> S (in hypophosphatasia; 4% of activity).
/FTId=VAR 011085.
249 G -> V (in hypophosphatasia; partial loss of activity).
IFTId=VAR 013988.
263 H -> Y (common polymorphism). lFTld=VAR
006161.
289 L -> F (in hypophosphatasia). /FTId=VAR
006162.
291 E -> K (in hypophosphatasia; moderate;
8% of activity).
/FTId=VAR 013989.
62 M -> L (in hypophosphatasia; moderate;
27% of activity).
/FTId=VAR 006148.
294 D -> A (in hypophosphatasia). /FTId=VAR
006163.
294 D -> Y (in hypophosphatasia). /FTId=VAR
013990.
306 D -> V (in hypophosphatasia). /FTId=VAR
006164.
326 G -> R (in hypophosphatasia; in a patient carrying also lys-291). /FTId=VAR 013991.
327 F -> G (in hypophosphatasia; requires 2 nucleotides substitutions). /FTId=VAR 013992.
327 F -> L (in hypophosphatasia; childhood).
/FTId=VAR 006165.
334 G -> D (in hypophosphatasia). /FTId=VAR
006166.
348 A -> T (in hypophosphatasia). /FTId=VAR
011086.
378 D -> V (in hypophosphatasia; loss of activity).
/FTId=VAR 006167.
381 H -> R (in hypophosphatasia). /FTId=VAR
O 11087.
63 G -> V (in hypophosphatasia; loss of activity).
IFTId=VAR 013974.
382 V -> I (in hypophosphatasia). IFTId=VAR
006168.
391 R -> C (in hypophosphatasia; moderate;
10% of activity).
/FTId=VAR 013993.
399 A -> S (in hypophosphatasia). IFTId=VAR
013994.
406 D -> G (in hypophosphatasia; 15% of activity).
/FTId=VAR Ol 1088.
423 V -> A (in hypophosphatasia; l6% of activity).
/FTId=VAR 013995.
426 G -> C (in hypophosphatasia; infantile;
partial loss of activity).IFTId=VAR 011089.
436 Y -> H (in hypophosphatasia). IFTId=VAR
006169.
445 S -> P (in hypophosphatasia; severe;
2% of activity).
lFTId=VAR 013996.
450 R -> C (in hypophosphatasia; severe;
4% of activity).
/FTId=VAR 013997.
450 R -> H (in hypophosphatasia). /FTId=VAR
011090.
71 R -> C (in hypophosphatasia). /FTId=VAR
006149.
456 G -> R (in hypophosphatasia; loss of activity).
/FTId=VAR 011091.
459 V -> M (in hypophosphatasia; infantile).
/FTId=VAR 013998.
473 G -> S (in hypophosphatasia). /FTId=VAR
013999.
476 E -> K (in hypophosphatasia). /FTId=VAR
006170.
478 N -> I (in hypophosphatasia; 9% of activity).
/FTId=VAR 011092.
489 C -> S (in hypophosphatasia; 9% of activity).
/FTId=VAR 011093.
490 I -> F (in hypophosphatasia; odonto;
partial loss of activity). /FTId=VAR 014000.
491 G -> R (in hypophosphatasia). IFTId=VAR
522 V -> A. iFTId=VAR 011094.
29 W -> A
71 R -> H (in hypophosphatasia). /FTId=VAR
104 N -> K
71 R -> P (in hypophosphatasia). /FTId=VAR
006150.
75 I G -> S (in hypophosphatasia; severe; 3.5% of activity).
/FTId=VAR 013976.
Protein Alkaline phosphatase, tissue-nonspecific isozyme precursor localization is believed to be attached to the membrane by a GPI-anchor.
The following GO Annotations) apply to the previously known protein. The following annotations) were found: skeletal development; ossification; metabolism, which are annotations) related to Biological Process; magnesium binding; alkaline phosphatase;
hydrolase, which are annotations) related to Molecular Function; and integral membrane protein, which are annotations) related to Cellular Component.
The GO assignment relies on information from one or more of the SwissProt/TremBl Protein knowledgebase, available from <http://www.expasy.ch/sprot/>; or Locuslink, available from <http://www.ncbi.nlm.nih.gov/projects/LocusLink/>.
As noted above, cluster HSAPHOL features 7 transcript(s), which were listed in Table 1 above. These transcripts) encode for proteins) which are variants) of protein Alkaline phosphatase, tissue-nonspecific isozyme precursor. A description of each variant protein according to the present invention is now provided.
Variant protein HSAPHOL P2 according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) HSAPHOL T4. An alignment is given to the known protein (Alkaline phosphatase, tissue-nonspecific isozyme precursor) at the end of the application. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between HSAPHOL P2 and AAH21289 (SEQ ID NO: 36):
l.An isolated chimeric polypeptide encoding for HSAPHOL P2, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PHSGPAAAFIRRRGWWPGPRCA corresponding to amino acids 1 - 22 of HSAPI-iOL P2, second amino acid sequence being at least 90 % homologous to PATPRPLSWLRAPTRLCLDGPSPVLCA corresponding to amino acids 1 - 27 of AAH21289, which also corresponds to amino acids 23 - 49 of HSAPHOL P2, and a third amino acid sequence being at least 90 % homologous to EKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFLGDGMGVSTVTAARILKGQL
HHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLCGVKANEGTVGVSAAT
MPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDVEYESDEKARGTRLD
GLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEPGDMQYELNRNNVT
DPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALHEAVEMDRAIGQAG
SLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYK
VVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQN
YVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 83 - 586 of AAH21289, which also corresponds to amino acids 50 -553 of HSAPHOL P2, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
2.An isolated polypeptide encoding for a head of HSAPHOL P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence PHSGPAAAFIRRRGWWPGPRCA of HSAPHOL P2.
3.An isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P2, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise AE, having a structure as follows: a sequence starting from any of amino acid numbers 49-x to 49; and ending at any of amino acid numbers SO+ ((n-2) - x), in which x varies from 0 to rr2.
Comparison report between HSAPHOL P2 and PPBT HUMAN
1.An isolated chimeric polypeptide encoding for HSAPHOL-P2, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence PHSGPAAAFIRRRGWWPGPRCAPATPRPLSWLRAPTRLCLDGPSPVLCA
corresponding to amino acids 1 - 49 of HSAPHOL P2, second amino acid sequence being at least 90 % homolo~us to EKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFLGDGMGVSTVTAARILKGQL
HHNFGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLCGVKANEGTVGVSAAT
ERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAAYAHSADRDWYSDNE
MPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDVEYESDEKARGTRLD
GLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEPGDMQYELNRNNVT
DPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALHEAVEMDRAIGQAG
SLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYK
VVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQN
YVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 21 - 524 of PPBT HUMAN, which also corresponds to amino acids 50 -553 of HSAPHOL P2, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
2.An isolated polypeptide encoding for a head of HSAPHOL P2, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence PHSGPAAAFIRRRGWWPGPRCAPATPRPLSWLRAPTRLCLDGPSPVLCA of HSAPHOL P2.
3.An isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P2, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about SO amino acids in length, wherein at least two amino acids comprise AE, having a structure as follows: a sequence starting from any of amino acid numbers 49-x to 49; and ending at any of amino acid numbers 50+ ((rr2) - x), in which x varies from 0 to rr2.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
membrane. The protein localization is believed to be membrane because although it is a partial protein, because both trans-membrane region prediction programs predict that this protein has a trans-membrane region, and similarity to known proteins suggests a GPI
anchor.Variant protein HSAPHOL P2 also has the following non-silent SNPs (Single Nucleotide Polymorphisms) as listed in Table 5, (given according to their positions) on the amino acid sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL P2 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 5 -Amino acid mutations a- "., "~~.F~. v~'X.. ,: . ,1' " ,. iT'~
~~ Y ~~. ., ,ES ~ W ' a ' non s : n-ammo° cid-;- Alte - atme~ mo. am . Preu~ousl known S .; "
SNP o~szt ; ~(. o , a gin, am d(s)." -; P~ y~ : ,~.
z ~a ~' ~ x~~ s~s~.~.' , 'i. -y "°,~,si~~a. a ,;icY & ". . t r i.., ~f F. Y
-. ~~,..'s. ~"a " c w:s~F.,~. ~t ~,~'r' .sr~m &a...
153 N -> S Yes 172 Q -> No 551 V -> A No 206 A -> No 272 R -> No 292 Y -> H Yes 342 V -> No 344 V -> No 354 K -> No 354 K -> Q No 380 E -> No Variant protein HSAPHOL P2 is encoded by the following transcript(s): HSAPHOL
T4, for which the sequences) is/are given at the end of the application. The coding portion of transcript HSAPHOL T4 is shown in bold; this coding portion starts at position 1 and ends at position 1659. The transcript also has the following SNPs as listed in Table 6 (given according to their position on the nucleotide sequence, with the alternative nucleic acid listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL P2 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 6 - Nucleic acid SNPs ;~.: ~ r _. ~",.,~ ,. . ~ :.==.
~~ ~-_ -~~ ~Previousl"' ~ kno n-SNP? . .. . ~.
SNP os~frop on nucleotide : ::Alternarivegnucleic acid~~. w _ ~.~p ,r'',~~r. a~~. ,:i >a. ~ »~a~-"=..,Fz ..~'r~ t~a°-~...~~.°
~~a'"'.,..'c~.r'''~'"~' ~r~~rri =...~..~-.-c ..~~.",~ ,.°. °cx.., iwm~.,~.-_..,:..~~;a.n~-~~~y ~...-.~'ss._. .y° a...,~ ~x ,_... ~..~F,-x:.~a.
417 C -> T Yes 458 A -> G Yes 1140 G -> No 1509 C -> T Yes 1629 G -> T Yes 1652 T -> C No 1727 C -> T Yes 1788 G -> A Yes 1895 A -> C Yes 2050 C -> T Yes 2095 A -> G Yes 2240 G -> No 516 G -> No 2347 -> A No 2364 T -> G No 617 C -> No 815 G -> No 874 T -> C Yes 1026 G -> No 1032 G -> No 1060 A -> No 1060 A -> C No Variant protein HSAPHOL P3 according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) HS APHOL T5. An alignment is given to the known protein (Alkaline phosphatase, tissue-nonspecific isozyme precursor) at the end of the application. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between HSAPHOL P3 and AAH21289:
1.An isolated chimeric polypeptide encoding for HSAPHOL P3, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVP
corresponding to amino acids 63 - 82 of AAH21289, which also corresponds to amino acids 1 -20 of HSAPHOL P3, and a second amino acid sequence being at least 90 % homologous to GMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYL
CGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSA
AYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTD
VEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFE
PGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQAL
HEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKK
PFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKG
PMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSV
LF corresponding to amino acids 123 - 586 of AAH21289, which also corresponds to amino acids 21 - 484 of HSAPHOL P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P3, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise PG, having a structure as follows: a sequence starting from any of amino acid numbers 20-x to 20; and ending at any of amino acid numbers 21+ ((m2) - x), in which x varies from 0 to n-2.
Comparison report between I-ISAPI-IOL P3 and PPBT HUMAN
l .An isolated chimeric polypeptide encoding for HSAPHOL P3, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVP
corresponding to amino acids 1 - 20 of PPBT HUMAN, which also corresponds to amino acids 1 -20 of 1-ISAPI-IOL_P3, and a second amino acid sequence being at least 90 %
homologous to GMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYL
CGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSA
AYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTD
VEYESDE.KARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFE
PGDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQAL
HEAVEMDRAIGQAGSLTSSEDTLTVVTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKK
PFTAILYGNGPGYKVVGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKG
PMAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSV
LF corresponding to amino acids 61 - 524 of PPBT_HUMAN, which also corresponds to amino acids 21 - 484 of HSAPHOL P3, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated chimeric polypeptide encoding for an edge portion of HSAPHOL P3, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise PG, having a structure as follows: a sequence starting from any of amino acid numbers 20-x to 20; and ending at any of amino acid numbers 21+ ((n-2) - x), in which x varies from 0 to rr2.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
membrane. The protein localization is believed to be membrane because of manual inspection of known protein localization and/or gene structure, and/or similarity to known proteins..
Variant protein HSAPHOL P3 also has the following non-silent SNPs (Single Nucleotide Polymorphisms) as listed in Table 7, (given according to the it positions) on the amino acid sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL P3 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 7 - Amino acid mzrtatiorrs ~,> :- ~s~:~..
. ~.. ,h~u.- . ~"~.~,. ..
t m. s, ax.r. .r id '~Alteriiatiue .,.-. ~ . , w~ .
SNI'= ositton s on~amtno ac 1? ~, ( ) ~ r am no,.aqid~.s- ~~'~emousl. ~known~SNP~ ~.~
c- ~~_ ~
_ . - x: 3 ,;x ..c r . ;. .~~.:. . - Wit... ,.;~, ; t .~ ' ~r w ~i_..H ~ ~ - .~~ s_ :_"'e' g'.n~ ~::;~,s~";f ,.~;. :W, ~ .:;;... C: ._.~ .a, -~r."~ .. P . ~-.a -.:., .:... R ~ ..,..~,~ -, m x--: _-~..~ .:~~"- ,x ~~ , ;5..~".<
use ~iienee =- su ~ ~ y~:v~ ~~~ _. ~ :-~~.~ y--., ~.'as.'"'.. w- ..,m'~.'..e:". .,~.~ . s~ ~ i,. ~,x ~"",..:. .,a~,s~",:a ~.,ir. r.,~~,~ ,~",; .. !i~ .. ~',~,..~.~1~.
~ 103 Q No 137 A -> No 84 N -> S Yes 1 -> No 203 R -> No 223 Y -> H Yes 273 V -> No 275 V -> No 285 K -> No 285 K -> Q No 311 E -> No 482 V -> A No Variant protein HSAPHOL P3 is encoded by the following transcript(s): HSAPHOL
T5, for which the sequences) is/are given at the end of the application. The coding portion of transcript HSAPHOL TS is shown in bold; this coding portion starts at position 253 and ends at position 1704. The transcript also has the following SNPs as listed in Table 8 (given according 10 to their position on the nucleotide sequence, with the alternative nucleic acid listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL P3 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 8 - Nucleic acid SNPs SNP position on nucleotideAlternative nucleic Previously.known acid SNP?
se uence. :~ ~ ~ ~,~ ~
q r ~~k -r ~.~ ~..~~:~- ~ : ~ a ,: ~
..._so - ..__. _ .. . ~ e_ 179 G -> C No 231 A -> No 1071 G -> No 1077 G -> No 1105 A -> No 1105 A -> C No 1185 G -> No 1554 C -> T Yes 1674 G -> T Yes 1697 T -> C No 1772 C -> T Yes 1833 G -> A Yes 232 A -> T No 1940 A -> C Yes 2095 C -> T Yes 2140 A -> G Yes 2285 G -> No 2392 -> A No 2409 T -> G No 281 T -> No 462 C -> T Yes 503 A -> G Yes 561 G -> No 662 C -> No 860 G -> No 919 T -> C Yes Variant protein HSAPHOL P4 according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) HSAPHOL_T6. An alignment is given to the known protein (Alkaline phosphatase, tissue-nonspecific isozyme precursor) at the end of the application. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between HSAPHOL P4 and AAH21289:
l.An isolated chimerie polypeptide encoding for HSAPHOL P4, comprising a first amino acid sequence being at least 90 % homologous to MGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLC
GVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAA
YAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDV
EYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEP
GDMQYELNRNNVTDPSLSEMVVVAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALH
EAVEMDRAIGQAGSLTSSEDTLTWTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKP
FTAILYGNGPGYKV VGGERENV SMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGP
MAHLLHGVHEQNYVPHVMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVL
F corresponding to amino acids 124 - 586 of AAH21289, which also corresponds to amino acids 1 - 463 of HSAPHOL P4.
Comparison report between HSAPHOL P4 and PPBT HUMAN
l .An isolated chimeric polypeptide encoding for HSAPHOL P4, comprising a first amino acid sequence being at least 90 % homologous to MGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTATAYLC
GVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHATPSAA
YAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPKNKTDV
EYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLLGLFEP
GDMQYELNRNNVTDPSLSEMVWAIQILRKNPKGFFLLVEGGRIDHGHHEGKAKQALH
EAVEMDRAIGQAGSLTSSEDTLTWTADHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKP
FTAILYGNGPGYKWGGERENVSMVDYAHNNYQAQSAVPLRHETHGGEDVAVFSKGP
MAHLLHGVHEQNYVPH VMAYAACIGANLGHCAPASSAGSLAAGPLLLALALYPLSVL
F corresponding to amino acids 62 - 524 of PPBT HUMAN, which also corresponds to amino acids 1 - 463 of HSAPHOL P4.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
membrane. The protein localization is believed to be membrane because only one of the two trans-membrane region prediction programs (Tmpred: l, Tmhmm: 0) has predicted that this protein has a trans-membrane region, but similarity to known proteins suggests a GPI anchor. In addition both signa~peptide prediction programs predict that this protein is a non-secreted protein..
Variant protein HSAPHOL P4 also has the following norrsilent SNPs (Single Nucleotide Polymorphisms) as listed in Table 9, (given according to their positions) on the amino acid 1 S sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL_P4 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 9 - Amino acid mutations t f.,., ~. ~C~~~' 7F4'n -.- ",.~t9 ~f :.,'" "~ , b ;.: a,~-...:. c. .~
~.,~ ~ ~a"~,....,.xm,". . .~..~,Eea$,.".. ~:a~ <ir~~".. i ~~
a~ s ~.x."." ~'.xu,~.~y";. ,-.~.
.. ~a .. c"=, ' ~ ~' . ~1~~'~m~tb~~.Cl~.-A,~ x:~a .~: a . re~~ously~ o o~ SlslF~
p , ( , ~ . ~~a ~ ~ t -;x x - ~W ~~~w ~..~,... ~.~tetatye4 a~n~no baGxd ;s~ .
-~~~r~ ~. a "' ~'. ,~ ~ ~,, ~ ~ as sequence= ~c ~
W ~ a ~,.. .~ '.'~~.... ~ ~'it-w . .:c~:~''~'~'"~~'~r~~~~x~~ x ~ ~s~, .,~ No ~
116 A ->
182 R -> No 82 Q -> No 202 Y -> H Yes 252 V -> No 254 V -> No 264 K -> No 264 K -> Q No 290 E -> No 461 V -> A No 63 J N -> S Yes Variant protein HSAPHOL P4 is encoded by the following transcript(s):
HSAPHOL_T6, for which the sequences) is/are given at the end of the application. The coding portion of transcript HSAPHOL-T6 is shown in bold; this coding portion starts at position 215 and ends at position 1603. The transcript also has the following SNPs as listed in Table 10 (given according to their position on the nucleotide sequence, with the alternative nucleic acid listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL P4 sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table 10 - Nucleic acid SNPs ,w r SNP ' ~h n~ ~. .~~, .,, ~.~ , . ~z, ... :~.~ ~ >
pos ~ o ~.o~ nueleQ.t~de . Alte -rnah~~-nu~'':~
:~~:~ ~,; ~ c exc. acrd: ee ;m zsl kn - wn -~ Y . o SN m .~
w -.~~. :m-~.~ ~ ~ =-. se ue c . , ~ o~ ~v ,.
Y 3:
~,~.
z ~..: ..a ~~ F ~ ::~.. ,x.s , ~;zs a ".,. "paYy"
~~,',.~r...m;.: . r... . ~ w".,: ' ~ .v;-.~'.. <.~'',~ ~:.~~=
361 C - j T Yes 402 A -> G Yes 1084 G _>
No 1453 C -> T
Yes 1573 G -> T
Yes 1596 T -> C No 1671 C -> T
Yes 1732 G -> A
Yes 1839 A -> C Yes 1994 C -> T Yes 2039 A -> G Yes 2184 G _>
No 460 G -> No 2291 -> A No 2308 T -> G No 561 C -> No 759 G -> No 818 T -> C Yes 970 G -> No 976 G -> No 1004 A -> No 1004 A -> C No Variant protein HSAPHOL PS according to the present invention has an amino acid sequence as given at the end of the application; it is encoded by transcripts) HSAPHOL_T7. An alignment is given to the known protein (Alkaline phosphatase, tissue-nonspecific isozyme precursor) at the end of the application. One or more alignments to one or more previously published protein sequences are given at the end of the application. A brief description of the relationship of the variant protein according to the present invention to each such aligned protein is as follows:
Comparison report between HSAPHOL PS and AAH21289:
l.An isolated chimeric polypeptide encoding for HSAPHOL P5, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGVSTVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
GLFEPGDMQYELNRNNVTDPSLSEMV V VAIQILRKNPKGFFLLVEGGRIDHGHHEGKA
KQALHEAVEM corresponding to amino acids 63 - 417 of AAH21289, which also corresponds to amino acids 1 - 355 of HSAPHOL PS, and a second amino acid sequence being at least 90 homologous to DHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVD
YAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIG
ANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 440 - 586 of AAH21289, which also corresponds to amino acids 356 - 502 of HSAPHOL P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated chimeric polypeptide encoding for an edge portion of I-ISAPHOL-P5, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise MD, having a structure as follows: a sequence starting from any of amino acid numbers 355-x to 355; and ending at any of amino acid numbers 356+ ((n-2) - x), in which x varies. from 0 to n-2.
Comparison report between HSAPHOL PS and PPBT HUMAN
l.An isolated chimeric polypeptide encoding for HSAPHOL P5, comprising a first amino acid sequence being at least 90 % homologous to MISPFLVLAIGTCLTNSLVPEKEKDPKYWRDQAQETLKYALELQKLNTNVAKNVIMFL
GDGMGV STVTAARILKGQLHHNPGEETRLEMDKFPFVALSKTYNTNAQVPDSAGTAT
AYLCGVKANEGTVGVSAATERSRCNTTQGNEVTSILRWAKDAGKSVGIVTTTRVNHA
TPSAAYAHSADRDWYSDNEMPPEALSQGCKDIAYQLMHNIRDIDVIMGGGRKYMYPK
NKTDVEYESDEKARGTRLDGLDLVDTWKSFKPRYKHSHFIWNRTELLTLDPHNVDYLL
GLFEPGDMQYELNRNNVTDPSLSEMVWAIQILRKNPKGFFLLVEGGRIDHGHHEGKA
KQALHEAVEM corresponding to amino acids 1 - 355 of PPBT HUMAN, which also corresponds to amino acids 1 - 355 of HSAPHOL P5, and a second amino acid sequence being at least 90 % homologous to DHSHVFTFGGYTPRGNSIFGLAPMLSDTDKKPFTAILYGNGPGYKVVGGERENVSMVD
YAHNNYQAQSAVPLRHETHGGEDVAVFSKGPMAHLLHGVHEQNYVPHVMAYAACIG
ANLGHCAPASSAGSLAAGPLLLALALYPLSVLF corresponding to amino acids 377 - 524 of PPBT HUMAN, which also corresponds to amino acids 356 - 502 of HSAPHOL_P5, wherein said first and second amino acid sequences are contiguous and in a sequential order.
2.An isolated chimeric polypeptide encoding for an edge portion of HSAPHOL_P5, comprising a polypeptide having a length "n", wherein n is at least about 10 amino acids in length, optionally at least about 20 amino acids in length, preferably at least about 30 amino acids in length, more preferably at least about 40 amino acids in length and most preferably at least about 50 amino acids in length, wherein at least two amino acids comprise MD, having a structure as follows: a sequence starting from any of amino acid numbers 355-x to 355; and ending at any of amino acid numbers 356+ ((n-2) - x), in which x varies from 0 to rr2.
The location of the variant protein was determined according to results from a number of different software programs and analyses, including analyses from SignalP and other specialized programs. The variant protein is believed to be located as follows with regard to the cell:
membrane. The protein localization is believed to be membrane because of manual inspection of known protein localization and/or gene structure and/or similarity to known protein..
Variant protein HSAPHOL PS also has the following norrsilent SNPs (Single Nucleotide Polymorphisms) as listed in Table I 1, (given according to their positions) on the amino acid sequence, with the alternative amino acids) listed; the last column indicates whether the SNP is known or not; the presence of known SNPs in variant protein HSAPHOL PS
sequence provides support for the deduced sequence of this variant protein according to the present invention).
Table ll -Amino acid mutations »- ., .~W x - ~~.W~
~S : ° ositso s, ~=on~amrno. aci .
P ~(.,~ ..,~ d~ Alternative arriino acrd ~ Prevro - -1:
~~~ r. ~ ~~ .; ~,~, , ~ a .uS y 1C110Wn S' ~4.~
.r ~ri ~..~'"~ A*."~
~ x~ ~ ° :~: ;~, ~: ~ m =~ :~: ~'.
124 ~ N -> S Yes 143 Q -> No 500 V -> A No 10 I -> No 1 ~~ A -> No 243 R -> No 263 Y -> H Yes 313 V -> No 315 V -> No 325 K -> No 325 K -> Q No 351 E -> No IS
DEMANDE OU BREVET VOLUMINEUX
LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND
PLUS D'UN TOME.
NOTE : Pour les tomes additionels, veuillez contacter le Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS
THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE THAN ONE
VOLUME
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Claims (52)
1. An isolated polynucleotide comprising a polynucleotide having a sequence selected from the group consisting of: R11723_PE_1_T15, R11723_PEA_1_T17, R11723_PEA_1_T19, R11723_PEA_1_T20, R11723_PEA_1_T5, or R11723_PEA_1_T6.
2. An isolated polynucleotide comprising a node having a sequence selected from the group consisting of:
3. An isolated polypeptide comprising a polypeptide having a sequence selected from the group consisting of: R11723_PEA_1_P2, R11723_PEA_1_P6, R11723_PEA_1_P7, R11723_PEA_1_P13, or R11723_PEA_1_P10.
4. An isolated chimeric polypeptide encoding for R11723_PEA_1_P6, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAGSPCRGLAPGREEQRALHKAGAVGGGVR
corresponding to amino acids 1 - 110 of R11723_PEA_1_P6, and a second amino acid sequence being at least 90 % homologous to MYAQALLVVGVLQRQAAAQHLHEHPPKLLRGHRVQERVDDRAEVEKRLREGEEDHV
RPEVGPRPVVLGFGRSHDPPNLVGHPAYGQCHNNQPWADTSRRERQRKEKHSMRTQ
corresponding to amino acids 1 - 112 of Q8IXM0, which also corresponds to amino acids 111 -222 of R11723_PEA_1_P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSAGIMYRKSCASSAACLIASAGSPCRGLAPGREEQRALHKAGAVGGGVR
corresponding to amino acids 1 - 110 of R11723_PEA_1_P6, and a second amino acid sequence being at least 90 % homologous to MYAQALLVVGVLQRQAAAQHLHEHPPKLLRGHRVQERVDDRAEVEKRLREGEEDHV
RPEVGPRPVVLGFGRSHDPPNLVGHPAYGQCHNNQPWADTSRRERQRKEKHSMRTQ
corresponding to amino acids 1 - 112 of Q8IXM0, which also corresponds to amino acids 111 -222 of R11723_PEA_1_P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
5. An isolated polypeptide encoding for a head of R11723_PEA_1_P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAGSPCRGLAPGREEQRALHKAGAVGGGVR of 811723_PEA_1_P6.
homologous to the sequence MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAGSPCRGLAPGREEQRALHKAGAVGGGVR of 811723_PEA_1_P6.
6. An isolated chimeric polypeptide encoding for R11723_PEA_1_P6, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 1 - 83 of Q96AC2, which also corresponds to amino acids 1 - 83 of R11723_PEA_1_P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ corresponding to amino acids 84 - 222 of R11723_PEA_1_P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 1 - 83 of Q96AC2, which also corresponds to amino acids 1 - 83 of R11723_PEA_1_P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ corresponding to amino acids 84 - 222 of R11723_PEA_1_P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
7. An isolated polypeptide encoding for a tail of R11723_PEA_1_P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKNSMRTQ in R11723_PEA-1_P6.
homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKNSMRTQ in R11723_PEA-1_P6.
8. An isolated chimeric polypeptide encoding for R11723_PEA_1_P6, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 1 - 83 of Q8N2G4, which also corresponds to amino acids 1 - 83 of R11723_PEA_1_P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ corresponding to amino acids 84 - 222 of R11723_PEA_1_P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 1 - 83 of Q8N2G4, which also corresponds to amino acids 1 - 83 of R11723_PEA_1_P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ corresponding to amino acids 84 - 222 of R11723_PEA_1_P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
9. An isolated polypeptide encoding for a tail of R11723_PEA_1_P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPWLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ in R11723_PEA_1_P6.
homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPWLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ in R11723_PEA_1_P6.
10. An isolated chimeric polypeptide encoding for R11723_PEA_1_P6, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 24 - 106 of BAC85518, which also corresponds to amino acids 1 - 83 of R11723_PEA_1_P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ corresponding to amino acids 84 - 222 of R11723_PEA_1_P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSAGIMYRKSCASSAACLIASAG corresponding to amino acids 24 - 106 of BAC85518, which also corresponds to amino acids 1 - 83 of R11723_PEA_1_P6, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ corresponding to amino acids 84 - 222 of R11723_PEA_1_P6, wherein said first and second amino acid sequences are contiguous and in a sequential order.
11. An isolated polypeptide encoding for a tail of R11723_PEA_1_P6, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ in R11723-PEA_1_P6.
homologous to the sequence SPCRGLAPGREEQRALHKAGAVGGGVRMYAQALLVVGVLQRQAAAQHLHEHPPKLL
RGHRVQERVDDRAEVEKRLREGEEDHVRPEVGPRPVVLGFGRSHDPPNLVGHPAYGQ
CHNNQPWADTSRRERQRKEKHSMRTQ in R11723-PEA_1_P6.
12. An isolated chimeric polypeptide encoding for R11723_PEA_1_P7, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAG corresponding to amino acids 1 - 64 of Q96AC2, which also corresponds to amino acids 1 - 64 of R11723_PEA_1_P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of R11723_PEA_1_P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSAG corresponding to amino acids 1 - 64 of Q96AC2, which also corresponds to amino acids 1 - 64 of R11723_PEA_1_P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of R11723_PEA_1_P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
13. An isolated polypeptide encoding for a tail of R11723_PEA_1_P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in R11723_PEA_1_P7.
homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in R11723_PEA_1_P7.
14. An isolated chimeric polypeptide encoding for R11723_PEA_1_P7, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAG corresponding to amino acids 1 - 64 of Q8N2G4, which also corresponds to amino acids 1 - 64 of R11723_PEA_1_P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of R11723_PEA_1_P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSAG corresponding to amino acids 1 - 64 of Q8N2G4, which also corresponds to amino acids 1 - 64 of R11723_PEA_1_P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of R11723_PEA_1_P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
15. An isolated polypeptide encoding for a tail of R11723_PEA_1_P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in R11723_PEA_1_P7.
homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in R11723_PEA_1_P7.
16. An isolated chimeric polypeptide encoding for R21723_PEA_1_P7, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWVLG corresponding to amino acids 1 - 5 of R11723_PEA_1_P7, second amino acid sequence being at least 90 % homologous to IAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEVMEQSAG
corresponding to amino acids 22 - 80 of BAC85273, which also corresponds to amino acids 6 -64 of R11723_PEA_1_P7, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of R11723_PEA_1_P7, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
corresponding to amino acids 22 - 80 of BAC85273, which also corresponds to amino acids 6 -64 of R11723_PEA_1_P7, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of R11723_PEA_1_P7, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
17. An isolated polypeptide encoding for a head of R11723_PEA_1_P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence MWVLG of R11723_PEA_1_P7.
homologous to the sequence MWVLG of R11723_PEA_1_P7.
18. An isolated polypeptide encoding for a tail of R11723_PEA_1_P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in R11723_PEA_1_P7.
homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in R11723_PEA_1_P7.
19. An isolated chimeric polypeptide encoding for R11723_PEA_1_P7, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSAG corresponding to amino acids 24 - 87 of BAC85518, which also corresponds to amino acids 1 - 64 of R11723_PEA_1_P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of R11723_PEA_1_P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSAG corresponding to amino acids 24 - 87 of BAC85518, which also corresponds to amino acids 1 - 64 of R11723_PEA_1_P7, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT corresponding to amino acids 65 - 93 of R11723_PEA_1_P7, wherein said first and second amino acid sequences are contiguous and in a sequential order.
20. An isolated polypeptide encoding for a tail of R11723_PEA_1_P7, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in R11723_PEA_1_P7.
homologous to the sequence SHCVTRLECSGTISAHCNLCLPGSNDHPT in R11723_PEA_1_P7.
21. An isolated chimeric polypeptide encoding for R11723_PEA_1_P13, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids 1 - 63 of Q96AC2, which also corresponds to amino acids 1 - 63 of R11723_PEA_1_P13, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DTKRTNTLLFEMRHFAKQLTT corresponding to amino acids 64 - 84 of R11723_PEA_1_P13, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSA corresponding to amino acids 1 - 63 of Q96AC2, which also corresponds to amino acids 1 - 63 of R11723_PEA_1_P13, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DTKRTNTLLFEMRHFAKQLTT corresponding to amino acids 64 - 84 of R11723_PEA_1_P13, wherein said first and second amino acid sequences are contiguous and in a sequential order.
22. An isolated polypeptide encoding for a tail of R11723_PEA_1_P13, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence DTKRTNTLLFEMRHFAKQLTT in R11723_PEA_1_P13.
homologous to the sequence DTKRTNTLLFEMRHFAKQLTT in R11723_PEA_1_P13.
23. An isolated chimeric polypeptide encoding for R11723_PEA_1_P10, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids 1 - 63 of Q96AC2, which also corresponds to amino acids 1 - 63 of R11723_PEA_1_P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of R11723_PEA_1_P10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSA corresponding to amino acids 1 - 63 of Q96AC2, which also corresponds to amino acids 1 - 63 of R11723_PEA_1_P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of R11723_PEA_1_P10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
24. An isolated polypeptide encoding for a tail of R11723_PEA_1_P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in R11723_PEA_1_P10.
homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in R11723_PEA_1_P10.
25. An isolated chimeric polypeptide encoding for R11723_PEA_1_P10, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids 1 - 63 of Q8N2G4, which also corresponds to amino acids 1 - 63 of R11723_PEA_1_P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of R11723_PEA_1_P10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSA corresponding to amino acids 1 - 63 of Q8N2G4, which also corresponds to amino acids 1 - 63 of R11723_PEA_1_P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of R11723_PEA_1_P10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
26. An isolated polypeptide encoding for a tail of R11723_PEA_1_P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence DRVSLCHEAGVQWIVNFSTLQPLPPRLK in R11723_PEA_1_P10.
homologous to the sequence DRVSLCHEAGVQWIVNFSTLQPLPPRLK in R11723_PEA_1_P10.
27. An isolated chimeric polypeptide encoding for R11723_PEA_1_P10, comprising a first amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence MWVLG corresponding to amino acids 1 - 5 of R11723_PEA_1_P10, second amino acid sequence being at least 90 % homologous to IAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEVMEQSA
corresponding to amino acids 22 - 79 of BAC8S273, which also corresponds to amino acids 6 -63 of R11723_PEA_1_P10, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of R11723_PEA_1_P10, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
corresponding to amino acids 22 - 79 of BAC8S273, which also corresponds to amino acids 6 -63 of R11723_PEA_1_P10, and a third amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90% and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of R11723_PEA_1_P10, wherein said first, second and third amino acid sequences are contiguous and in a sequential order.
28. An isolated polypeptide encoding for a head of R11723_PEA_1_P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence MWVLG of R11723_PEA_1_P10.
homologous to the sequence MWVLG of R11723_PEA_1_P10.
29. An isolated polypeptide encoding for a tail of R11723_PEA_1_P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in R11723_PEA_1_P10.
homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in R11723_PEA_1_P10.
30. An isolated chimeric polypeptide encoding for R11723_PEA_1_P10, comprising a first amino acid sequence being at least 90 % homologous to MWVLGIAATFCGLFLLPGFALQIQCYQCEEFQLNNDCSSPEFIVNCTVNVQDMCQKEV
MEQSA corresponding to amino acids 24 - 86 of BAC85518, which also corresponds to amino acids 1 - 63 of R11723_PEA_1_P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of R11723_PEA_1_P10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
MEQSA corresponding to amino acids 24 - 86 of BAC85518, which also corresponds to amino acids 1 - 63 of R11723_PEA_1_P10, and a second amino acid sequence being at least 70%, optionally at least 80%, preferably at least 85%, more preferably at least 90%
and most preferably at least 95% homologous to a polypeptide having the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK corresponding to amino acids 64 - 90 of R11723_PEA_1_P10, wherein said first and second amino acid sequences are contiguous and in a sequential order.
31. An isolated polypeptide encoding for a tail of R11723_PEA_1_P10, comprising a polypeptide being at least 70%, optionally at least about 80%, preferably at least about 85%, more preferably at least about 90% and most preferably at least about 95%
homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in R11723_PEA_1_P10.
homologous to the sequence DRVSLCHEAGVQWNNFSTLQPLPPRLK in R11723_PEA_1_P10.
32. An isolated oligonucleotide, comprising an amplicon selected from the group consisting of SEQ ID NOs: 975 or 978.
33. A primer pair, comprising a pair of isolated oligonucleotides capable of amplifying said amplicon of claim 32.
34. The primer pair of claim 33, comprising a pair of isolated oligonucleotides selected from the group consisting of: SEQ NOs 972 and 973; or 976 and 977.
35. An antibody capable of specifically binding to an epitope of an amino acid sequence of any of claims 3-31.
36. The antibody of claim 35, wherein said amino acid sequence comprises said tail of claims 4-31.
37. The antibody of claims 35 or 36, wherein said antibody is capable of differentiating between a splice variant having said epitope and a corresponding known protein PSEC.
38. A kit for detecting ovarian cancer, comprising a kit detecting overexpression of a splice variant according to any of the above claims.
39. The kit of claim 38, wherein said kit comprises a NAT-based technology.
40. The kit of claim 39, wherein said kit further comprises at least one primer pair capable of selectively hybridizing to a nucleic acid sequence according to claims 1 or 2.
41. The kit of claim 38, wherein said kit further comprises at least one oligonucleotide capable of selectively hybridizing to a nucleic acid sequence according to claims 1 or 2.
42. The kit of claim 38, wherein said kit comprises an antibody according to any of claims 35-37.
43. The kit of claim 42, wherein said kit further comprises at least one reagent for performing an ELISA or a Western blot.
44. A method for detecting ovarian cancer, comprising detecting overexpression of a splice variant according to any of the above claims.
45. The method of claim 44, wherein said detecting overexpression is performed with a NAT-based technology.
46. The method of claim 44, wherein said detecting overexpression is performed with an immunoassay.
47. The method of claim 46, wherein said immunoassay comprises an antibody according to any of the above claims.
48. A biomarker capable of detecting ovarian cancer, comprising any of the above nucleic acid sequences or a fragment thereof, or any of the above amino acid sequences or a fragment thereof.
49. A method for screening for ovarian cancer, comprising detecting ovarian cancer cells with a biomarker or an antibody or a method or assay according to any of the above claims.
50. A method for diagnosing ovarian cancer, comprising detecting ovarian cancer cells with a biomarker or an antibody or a method or assay according to any of the above claims.
51. A method for monitoring disease progression and/or treatment efficacy and/or relapse of ovarian cancer, comprising detecting ovarian cancer cells with a biomarker or an antibody or a method or assay according to any of the above claims.
52. A method of selecting a therapy for ovarian cancer, comprising detecting ovarian cancer cells with a biomarker or an antibody or a method or assay according to any of the above claims and selecting a therapy according to said detection.
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US62811204P | 2004-11-17 | 2004-11-17 | |
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US63055904P | 2004-11-26 | 2004-11-26 | |
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PCT/IB2005/002555 WO2005116850A2 (en) | 2004-01-27 | 2005-01-27 | Differential expression of markers in ovarian cancer |
US- | 2005-07-11 |
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CA2554703A1 true CA2554703A1 (en) | 2005-12-08 |
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CA002554703A Abandoned CA2554703A1 (en) | 2004-01-27 | 2005-01-27 | Differential expression of markers in ovarian cancer |
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EP (1) | EP1721257A2 (en) |
AU (1) | AU2005248530A1 (en) |
CA (1) | CA2554703A1 (en) |
WO (1) | WO2005116850A2 (en) |
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US8642031B2 (en) | 2006-11-02 | 2014-02-04 | Acceleron Pharma, Inc. | Antagonists of BMP9, BMP10, ALK1 and other ALK1 ligands, and uses thereof |
US10059756B2 (en) | 2006-11-02 | 2018-08-28 | Acceleron Pharma Inc. | Compositions comprising ALK1-ECD protein |
KR20170012582A (en) | 2006-11-02 | 2017-02-02 | 악셀레론 파마 인코포레이티드 | Alk1 receptor and ligand antagonists and uses thereof |
BRPI0911853A8 (en) | 2008-05-02 | 2018-03-06 | Acceleron Pharma Inc | compositions and methods for angiogenesis modulation and pericyte composition |
DK4209510T5 (en) | 2008-12-09 | 2024-07-22 | Hoffmann La Roche | ANTI-PD-L1 ANTIBODIES AND THEIR USE IN PROMOTING T CELL FUNCTION |
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US7022821B1 (en) * | 1998-02-20 | 2006-04-04 | O'brien Timothy J | Antibody kit for the detection of TADG-15 protein |
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- 2005-01-27 EP EP05780004A patent/EP1721257A2/en not_active Ceased
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- 2005-01-27 AU AU2005248530A patent/AU2005248530A1/en not_active Abandoned
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WO2005116850A2 (en) | 2005-12-08 |
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