[go: up one dir, main page]
More Web Proxy on the site http://driver.im/

WO2002037115A1 - Détection d'agents infectieux au moyen de mimiques d'antigènes - Google Patents

Détection d'agents infectieux au moyen de mimiques d'antigènes Download PDF

Info

Publication number
WO2002037115A1
WO2002037115A1 PCT/IT2000/000442 IT0000442W WO0237115A1 WO 2002037115 A1 WO2002037115 A1 WO 2002037115A1 IT 0000442 W IT0000442 W IT 0000442W WO 0237115 A1 WO0237115 A1 WO 0237115A1
Authority
WO
WIPO (PCT)
Prior art keywords
phage
hcv
collection
sera
clones
Prior art date
Application number
PCT/IT2000/000442
Other languages
English (en)
Inventor
Franco Felici
Nicola Gargano
Olga Minenkova
Paolo Monaci
Original Assignee
Kenton S.R.L.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kenton S.R.L. filed Critical Kenton S.R.L.
Priority to BR0017366-5A priority Critical patent/BR0017366A/pt
Priority to SK536-2003A priority patent/SK5362003A3/sk
Priority to CZ20031044A priority patent/CZ20031044A3/cs
Priority to PL00364832A priority patent/PL364832A1/xx
Priority to MXPA03003794A priority patent/MXPA03003794A/es
Priority to KR10-2003-7006153A priority patent/KR20030084895A/ko
Priority to CA002427602A priority patent/CA2427602A1/fr
Priority to EP00981607A priority patent/EP1332369A1/fr
Priority to CN00820000A priority patent/CN1455866A/zh
Priority to PCT/IT2000/000442 priority patent/WO2002037115A1/fr
Priority to AU2001218836A priority patent/AU2001218836A1/en
Priority to HU0302103A priority patent/HUP0302103A3/hu
Priority to JP2002539818A priority patent/JP2004513346A/ja
Publication of WO2002037115A1 publication Critical patent/WO2002037115A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
    • G01N33/5767—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis non-A, non-B hepatitis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/24011—Flaviviridae
    • C12N2770/24022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/24011—Flaviviridae
    • C12N2770/24211—Hepacivirus, e.g. hepatitis C virus, hepatitis G virus
    • C12N2770/24222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • the present invention relates to a diagnostic assay for detecting infectious agents, in particular viral agents, more in particular human Hepatitis C Virus (hereinafter briefly named HCV), anti-infectious agent antibody binding peptides useful in said assay, to processes for preparing said peptides and to kits for performing said assay.
  • HCV human Hepatitis C Virus
  • the present invention relates in particular to HCV, anti- HCV antibody binding peptides useful in diagnostic assay, processes for the preparation thereof and to kits for performing said assays. Background of the invention
  • Hepatitis C virus is the major etiological agent of parenterally transmitted, non-A, non-B hepatitis. This virus very often causes persistent infection and frequently leads to the development of chronic hepatitis and liver cirrhosis, constituting a major worldwide cause of chronic liver disease (Boyer N, Marcellin P, Pathogenesis, diagnosis and management of hepatitis C, J Hepatol 2000; 32:98-112).
  • Viral infection is diagnosed by detecting anti-HCV antibodies in the serum or by revealing viral RNA through nucleic acid amplification methods (Robbins DJ, Pasupuleti V, Cuan J, Chiang CS, Reverse transcriptase PCR quantitation of hepatitis C virus, Clin Lab Sci 2000 Winter; 13:23-30). These latter methods are very sensitive, but are expensive and prone to technical or laboratory error. In addition, reliability and specificity of the PCR technique are not standardized (Gretch DR, Diagnostic tests for hepatitis C, Hepatology 1997 Sep;26(3 Suppl 1):43S-47S)
  • the present invention relates to diagnosis of infectious diseases, such as viral infections, in particular to HCV infections, by means of detecting antibodies raised against the infectious agent, in particular anti-HCV antibodies.
  • US 5,985,542 assigned to Sumitomo Chemical Company, provides a kit for liver diseases, such as hepatitis C and alcoholic cirrh ⁇ sis7 which contains an antibody capable of recognizing cytochrome P450.
  • US 5,972,347 assigned to Baxter Aktiengeselleschaft, provides a composition for the treatment of HCV infection comprising inactivated HCV bound to neutralizing human HCV antibodies, wherein the antibodies are against at least one protein selected from the group consisting of HCV-core protein and NS3-protein.
  • US 5,800,982, assigned to Tonen discloses antigenic peptides capable of reacting specifically with antibodies directed against Group II of HCV.
  • JP 10019897, to Tonen provides a kit containing a peptide having amino acid sequence of at least five continuous amino acids constituting protein of a non-structure region 4A (NS4A) of HCV and peptide having amino acid sequence of at least five continuous amino acids constituting protein of a non- structure region 4B (NS4B) of HCV.
  • the contact of a foreign antigen with an organism activates a specific immune response.
  • the image of the antigen that is detected by the humoral response is defined by the epitopes recognized by the host antibodies. Assuming that the antibody binding site is the negative image of the epitope, a molecule that specifically binds the paratope should represent a positive image of the epitope. This line of thought suggests that, as far as humoral response is concerned, an antigen could be faithfully described by specific ligands that bind to the antigen- specific antibodies. Identifying these ligands would offer a way to detect the specific humoral response against an antigen, independently from whether or not that same antigen is known and/ or available.
  • ADAM-HCV EIA Enzymatic Immuno Assay
  • the size of the phage particle limits the concentration of peptide molecules that can be achieved in the assay.
  • Interference of serum antibodies against the phage capsid requires the addition of carrier phage in the assay mixture to sequester anti-phage antibodies.
  • the large-scale production of this, as any, biological reagent faces problems of microbiological contamination, reproducibility, quality control, purification and production costs.
  • a method for making a diagnosis of infectious diseases such as viral infections, in particular to hepatitis C, comprising identifying the binding specificity of the anti-antigen antibody molecules in the serum by the Antibody Detection by Antigen Mimics (ADAM) methodology, comprising screening phage libraries using sera from antigen-infected patients and non-infected individuals, identifying peptides binding antibodies (ligands) specifically associated with said antigen.
  • infectious diseases such as viral infections, in particular to hepatitis C
  • ADAM Antibody Detection by Antigen Mimics
  • the present invention relates to HCV infection.
  • said ligands are improved by in vitro maturation strategies.
  • said ligands are synthetic peptides.
  • said ligands are linked to a common core.
  • said ligands, together with said common core is MAP, according to the definition given hereinafter. It is also another object of the present invention a collection of
  • HCV-specific ligands obtainable by the process comprising: a) first panning a phage library on n positive sera to generate a first series of n phage pools; b) preparing n pool mixtures containing n-1 pools; c) selection affinity of each of n mixtures against the serum that generated the excluded pool of phage, to give a second series of n phage pools, and optionally d) additional panning each of the second series of n phage pools on a mix composed of all the n original sera, except that used for the first panning; e) immunoscreening of the resulting second series of n phage pools using a mixture of all the n original sera to give positive clones; f) testing the individual reactivity of all the positive clones with a panel of positive and negative sera by using an ordered array of said clones as phage- secreting colonies; g) generating replicas of said phage-secreting colonies; h) screening each replica for their re
  • YSREQLNKLFGIEVM YSREQLSKLFGIDTQ; KSREQLSKLHGVDTS; RSREQLSKLFGIDLT; - MWRTWLM THGIESW;
  • AEGEFRLGVRALRKALDPAK - AEGEFRLGVRALRKAPDPAK
  • AEGEFYSREQLNKLFGIDMTDPAK - AEGEFYSREQLNKMFGIETSDPAK;
  • AEGEFYSPEWLNKARGIDRSDPAK - AEGEFKSREQLSKLHGVDTSDPAK;
  • a preferred phage library of step a) is pVIII- 12aa.
  • the inserts of the preferred clones singled out from step k have the sequences
  • a phage library is generated in which the reacting clones, more preferably the best reacting clones, are partially mutagenized so that each amino acid of the clone sequence is independently substituted by any other amino acid.
  • said clones have the preferred insert sequence: SREQLNKLFGIEG.
  • the random sequence in the phage library, may be flanked by two cysteine residue. This aspect may apply in the general exploitation of the present invention and may not be limited to the case of HCV. It is an object of the present invention the use of the above collection for the preparation of a diagnostic assay for detecting infectious agents, such as viruses, in particular HCV in a subject suspected to be affected by said infectious agents, such as viruses, in particular HCV. It is an object of the present invention a kit for diagnostic purposes, comprising the above collection.
  • the immunogenic peptides obtainable, either in the form of a collection or as single peptide, from, the process herein disclosed. Said peptides are useful as immunogenic, therefore are useful for the preparation of vaccines, in particular against HCV. Conveniently, the peptides are in the form of the kit above mentioned.
  • the diagnostic assay we tested has an in-built upgrading capacity: ad hoc selections can be performed with those sera for which no response was detected.
  • a similar methodology might be used to identify a peptide panel which discriminates among different HCV genotypes, thus replacing expensive and labour-intensive PCR methods with a cheaper and faster EIA.
  • the present invention shall be described in further details also by means of examples and figures, wherein in the latter:
  • Figure 1 represents the characterization of clones derived from screening. The amino acid sequences of selected clones are reported in single letter code. Top and lower panels display sequences derived from the original or the secondary library, respectively. pVIII sequences flanking the foreign epitope are (NH2)AEGEF[foreign epitope] DPAK. Gray boxes indicate residues more frequently present at any given position in clones from original library. Residues contributing to the consensus sequence of the peptides from the secondary library are indicated in bold characters.
  • Figure 2 represents the identification of phage mimicking the same antigen determinant.
  • Antibodies affinity purified by positive serum C65 by clones PAl, PA3, PA8, PA 12 or P18 (listed in the first column) were tested in ELISA for their reactivity against the same phage (listed in the first row) .
  • Figure 3 represents the ELISA reactivity of pool derived from selection of library pVIIIA12. Phage pool derived from library panning library pVIIIA12 on positive sera C76 was tested in ELISA for its reactivity with positive sera C12, C13, C29, c40, C47, C65, C73, C74, C76, C83 and C85.
  • ELISA as described in Materials and Methods detected binding of peptides to antibodies present in human sera. Average values from two independent experiments were collected. Results are expressed as the ratio between the measure signal and the cut-off value (S/CO). Number of sera tested for each group of sera is indicated.
  • Figure 5 ADAM /HCV EIA on a panel of indeterminate sera.
  • the left-most column indicates the name of the HCV peptides tested, grouped according to their binding specificity.
  • the next four columns report the reactivities of the listed peptides with positive (c25 and rl5) and negative (r6 and rl3) control sera.
  • Each additional column reports the reactivities of the listed peptides with indeterminate sera.
  • Binding of antibodies to HCV peptides present in human sera was detected by ELISA as described in Material and Methods. Average values from two independent experiments have been determined.
  • FIG. 6 ADAM-HCV/ SIA on positive, negative and indeterminate sera.
  • the following ADAM-HCV peptides were grouped according to their binding specificity and immobilized onto nylon membrane to obtain 10 bands: ml909.2 and ml913.2 (A); ml901.31, m3322.3, m3362.3 (B); ml977.1 (C); m3551.3 (D); m3566.3 (E); m858, mF78 and mHl (F); mA12.1, mA12.2 and mA12.12 (G); mB11.17 (H); mG21.2 (I); ml929A3.1, ml929C3.4 and ml 929.21 (J).
  • An additional line containing purified human IgG was included as internal positive control. Binding of antibodies to HCV peptides present in human sera was detected as described in Material and Methods
  • MAP antigen peptides
  • Tarn, J.P. Synthetic peptide vaccine design: synthesis and properties of a high- density multiple antigenic peptide system. Proc. Natl. Acad. Sci. 85 (1988) 5409-5413)
  • MAP antigen peptides
  • Tarn, J.P. Synthetic peptide vaccine design: synthesis and properties of a high- density multiple antigenic peptide system. Proc. Natl. Acad. Sci. 85 (1988) 5409-5413
  • the removal of the phage scaffold allows a higher peptide concentration of immobilized ligand, which results in a higher sensitivity.
  • the panel of HCV- specific ligands obtainable according to the present invention includes peptides mimicking HCV NS3, an immunodominant HCV antigen.
  • this is the first time that a short peptide which mimics an immunodominant NS3 determinant is described.
  • Pepscan analysis of the NS3 protein revealed longer sequences which rarely reacted with positive sera (Khudyakov, Y., et al. 1995. Virology, 206:666-672).
  • Phage displayed random peptide libraries represent a powerful tool to identify ligands which are specifically recognized by anti-HCV serum antibodies. Phage-borne peptides have a broad mimicking potential, as they are able to mimic linear, conformational and even non proteinaceous epitopes (for a review see Felici F, Luzzago A, Monaci P, Nicosia A, Sollazzo M, Traboni C. Peptide and protein display on the surface of filamentous bacteriophage. Biotechnol Annu Rev. 1995; 1 : 149-83; Zwick MB, Shen J, Scott JK. Phage-displayed peptide libraries. Curr Opin Biotechnol.-.1998 Aug;9(4):427-36).
  • compositions comprising immunogens and vaccines according to the present invention are prepared conventionally as normally understood by those having ordinary experience in the art. For example, they may be prepared as described in EP 0 698 091. The following example further illustrate the invention.
  • Phage library pVIII-12aa was panned on 8 positive sera (C13,
  • pool pl3 ⁇ was panned on a mixture of sera C14, C27,
  • a phage library was generated in which the sequence of clone PA12 was partially mutagenized.
  • this "secondary" library named pVIIIA12
  • oligonucleotides were synthesized so that each amino acid of the SREQLNKLFGIEG sequence was independently substituted by any other amino acid: in theory, a substitution at each position would occur at a frequency of 20 percent.
  • a random residue was included at both sites of the foreign peptide sequence.
  • the pVHIA12 library was panned twice on 12 positive sera (C8, CIO, C12, C13, C22, C58, C60, C76, C83, C85, C141 arid C177).
  • phage pools ⁇ 76 ⁇ , pl41 and ⁇ l77 ⁇ (derived from selection using sera C76, C141 and C177, respectively) showed the highest and broadest reactivity with a panel of positive sera and were further analyzed ( Figure 3).
  • phage pools p76 ⁇ , p l41 ⁇ and pl77 ⁇ were immunoscreened by using sera C40, C141 and C177, respectively.
  • this analysis picked out several clones that were then individually tested for their reactivity with many different positive and negative sera by a filter-replica protocol, as detailed above. This final screening' singled out 51 clones specifically reacting with positive sera.
  • Phage libraries of various lengths were screened in which the random sequence was either completely random or flanked by two cysteine residues that constrain the conformation of the displayed peptide.
  • Phage pools exhibiting interesting reactivity profiles with positive sera were further analyzed. Assessing the reactivity of a large number of individual clones by replica screening singled out phage displaying a specific reactivity with positive sera.
  • HVR1 phage-borne peptides derived from this screening was analyzed for their reactivity with our panel of sera. Three peptides were identified with the highest and specific frequency of reactivity with positive sera (mF78, mHl and m858).
  • HCV-ligands were synthesized as octa-branching multiple antigen peptides (ADAM-HCV peptides).
  • ADAM-HCV peptides eight identical peptide sequences are linked through a lysine fork to a common core to form a multiple display similar to that of pVIII-fused peptides on the phage capsid.
  • pVIII sequences flanking the foreign epitope were shown to be relevant for the binding specificity of the corresponding peptide.
  • a mixture containing these 22 ADAM-HCV peptides was used to detect the presence of anti-HCV antibodies by an EIA (ADAM/ HCV EIA).
  • the ADAM-HCV mix of peptides was immobilized by passive coating at the bottom of a multi-well ELISA plate and incubated with 1 :40 diluted sera samples for 40 min. Human antibodies bound to the peptides were detected by incubating for 20 min with anti-human conjugate and measured through a chromogenic enzymatic reaction.
  • ADAM-HCV EIA efficiently discriminates between positive and negative sera.
  • ADAM-HCV strip immunoblot assay (ADAM-HCV /SIA).
  • ADAM-HCV peptides were covalently immobilized onto an activated nylon membrane to obtain a strip with ten bands.
  • Each line included different peptides with the same binding specificity, as detailed in the legend of Figure 6.
  • a control line containing purified human IgG was also included as internal positive control.
  • a selected number of samples from the panel of indeterminate sera were tested for their reactivity with immobilized antigens by incubating serum samples with the strip.
  • Anti-HCV antibodies captured by individual antigens were visualized by incubating the strips with anti-human enzyme-conjugate followed by a colorimetric enzymatic reaction. The reactivity of specimens with peptide bands was determined by visually comparing the intensity of each band with that of the internal positive control.
  • ADAM-HCV/ SIA revealed the reactivity of all the 8 positive sera tested to several different viral determinants mimicked by ADAM- HCV peptides. No reactivity was detected when 8 negative sera were tested ( Figure 6).
  • pVIII9aa pVIII9aa_cys
  • pVIII12aa pVIII15aa
  • pVIIIA12aa pVIII9aa (Felici et al., 1991)
  • pVIII12aa and pVIII15aa are three different libraries composed of random 9-mers
  • pVIII9aa_cys is a library in which the random nonapeptide is flanked by two cysteine residues (Luzzago et al., 1993). In this latter library, cysteines promote the formation of a disulfide bridge that constrains to some extent the conformation of the displayed peptide.
  • the pVIIIA12 library was constructed by synthesizing an oligonucleotide encoding the amino acid sequence SREQLNKLFGIEG.
  • Sera were also tested for the absence of antibodies to HBsAg and to HIV-l/HIV-2 by AUSAB EIA test (Abbott Labs, Chicago, IL) and by the third generation HIV- l/HIV-2 EIA test (Abbott Labs, South Pasadena, CA). Samples positive for the presence of anti-HCV antibodies, but negative for anti-HBsAg and anti-HIV antibodies, were included in this study as HCV-positive sera. Sera negative for the presence of antibodies against all the three antigens were included in this study as HCV- negative sera.
  • Phage supernatants were prepared from DH5 ⁇ -F' infected cells as previously described (Felici et al., 1991). Multi-well plates (Immunoplate Maxisorp, Nunc, Roskilde, Denmark) were coated overnight at 4°C with 200 ⁇ l of the anti-pill monoclonal antibody 57D1 (Dente et al., 1994) at a concentration of 1 ⁇ g of antibody/ml in 50 mM NaHC0 3 pH 9.6. After discarding coating solution, plates were incubated at 37°C for 60 min with ELISA blocking buffer (0.1% casein, 1% Triton-XlOO in PBS).
  • the p value referred to clones PA8 and PA 12 is the probability that the observed frequency distributions of reactivities of the positive and negative sera are statistically the same according to the ⁇ test.
  • Affinity-purified antibodies were tested in standard ELISA for their reactivity against phagotopes.
  • multi-well plates were coated with 100 ⁇ l/well of a solution of 1x10 H TU/ml CsCl-purified phage in 50 mM NaHC ⁇ 3 pH 9.6, overnight at 4°C. After washing with PBS/Tween, plates were incubated for 60 min at 37°C with blocking buffer. Afterwards, 100 ⁇ l of affinity-purified antibodies were added to each well and allowed to bind overnight at 4°C.
  • MAPs synthetic octabranching multiple antigen peptides
  • Multi-well plates (Immuno plate Maxisorp, Nunc, Roskilde, Denmark) were coated overnight at 4°C with a MAP solution at a concentration of 10 ⁇ g /ml in 50 m ' M NaHC ⁇ 3 pH 9.6. After discarding coating solution, plates were incubated at 37°C for 60 min with ELISA blocking buffer (0.1% casein, 1% Triton-XlOO in PBS). Plates were washed several times with PBS/0.05% Tween-20 (washing buffer). A 1:40 diluted human serum was added to each well and incubated 40 min 37°C. Plates were then washed with washing buffer, and a 1 :20.000 dilution of goat anti-human IgG HRP-conj.
  • HCV hepatitis C virus

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pathology (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • General Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Food Science & Technology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Virology (AREA)
  • Communicable Diseases (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé permettant d'effectuer le diagnostic d'un antigène. Ce procédé consiste à identifier la spécificité de liaison des molécules de l'anticorps anti-antigène dans le sérum au moyen de la méthode ADAM (détection d'un anticorps par mimiques d'antigènes). Ce procédé consiste à cribler des bibliothèques de phages au moyen de sérums tirés de patients infectés par des antigènes et des sujets non infectés et à identifier des anticorps se liant aux peptides (ligands) associés de manières spécifique audit antigène. Pour améliorer ledit procédé, on utilise des stratégies de maturation in vitro, on lie les ligands à un noyau commun, tel que MAP. Ledit procédé s'applique notamment au virus de l'hépatite C.
PCT/IT2000/000442 2000-11-03 2000-11-03 Détection d'agents infectieux au moyen de mimiques d'antigènes WO2002037115A1 (fr)

Priority Applications (13)

Application Number Priority Date Filing Date Title
BR0017366-5A BR0017366A (pt) 2000-11-03 2000-11-03 Detecção de agentes infecciosos usando imitadores de antìgeno
SK536-2003A SK5362003A3 (en) 2000-11-03 2000-11-03 Detection of infectious agents using antigen mimics
CZ20031044A CZ20031044A3 (cs) 2000-11-03 2000-11-03 Způsob diagnostikování infekčních agens s využitím antigenových mimetik
PL00364832A PL364832A1 (en) 2000-11-03 2000-11-03 Detection of infectious agents using antigen mimics
MXPA03003794A MXPA03003794A (es) 2000-11-03 2000-11-03 Deteccion de agentes infecciosos utilizando antigenos imitadores.
KR10-2003-7006153A KR20030084895A (ko) 2000-11-03 2000-11-03 항원 유사성을 이용하여 감염 인자를 검출하는 방법
CA002427602A CA2427602A1 (fr) 2000-11-03 2000-11-03 Detection d'agents infectieux au moyen de mimiques d'antigenes
EP00981607A EP1332369A1 (fr) 2000-11-03 2000-11-03 D tection d'agents infectieux au moyen de mimiques d'antig nes
CN00820000A CN1455866A (zh) 2000-11-03 2000-11-03 使用抗原模拟物的传染原的检测
PCT/IT2000/000442 WO2002037115A1 (fr) 2000-11-03 2000-11-03 Détection d'agents infectieux au moyen de mimiques d'antigènes
AU2001218836A AU2001218836A1 (en) 2000-11-03 2000-11-03 Detection of infectious agents using antigen mimics
HU0302103A HUP0302103A3 (en) 2000-11-03 2000-11-03 Detection of infectious agents using antigen mimics
JP2002539818A JP2004513346A (ja) 2000-11-03 2000-11-03 診断方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/IT2000/000442 WO2002037115A1 (fr) 2000-11-03 2000-11-03 Détection d'agents infectieux au moyen de mimiques d'antigènes

Publications (1)

Publication Number Publication Date
WO2002037115A1 true WO2002037115A1 (fr) 2002-05-10

Family

ID=11133576

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IT2000/000442 WO2002037115A1 (fr) 2000-11-03 2000-11-03 Détection d'agents infectieux au moyen de mimiques d'antigènes

Country Status (12)

Country Link
EP (1) EP1332369A1 (fr)
JP (1) JP2004513346A (fr)
KR (1) KR20030084895A (fr)
CN (1) CN1455866A (fr)
AU (1) AU2001218836A1 (fr)
BR (1) BR0017366A (fr)
CA (1) CA2427602A1 (fr)
HU (1) HUP0302103A3 (fr)
MX (1) MXPA03003794A (fr)
PL (1) PL364832A1 (fr)
SK (1) SK5362003A3 (fr)
WO (1) WO2002037115A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2108656A1 (fr) 2008-03-19 2009-10-14 Beninati, Concetta Fragments de protéines antigéniques de streptococcus pneumoniae
WO2009158682A2 (fr) * 2008-06-27 2009-12-30 Watkinson D Tobin Compositions et procédés pour diagnostiquer et traiter des troubles pathogènes

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0744467A2 (fr) * 1995-03-30 1996-11-27 Eisai Co., Ltd. Peptide multi-antigénique comprenant au moins deux peptides associés avec le virus de l'hépatite C
WO1999058561A1 (fr) * 1998-05-14 1999-11-18 Pasteur Merieux Serums & Vaccins Mimotopes du virus de l'hepatite c

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0744467A2 (fr) * 1995-03-30 1996-11-27 Eisai Co., Ltd. Peptide multi-antigénique comprenant au moins deux peptides associés avec le virus de l'hépatite C
WO1999058561A1 (fr) * 1998-05-14 1999-11-18 Pasteur Merieux Serums & Vaccins Mimotopes du virus de l'hepatite c

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
BARTOLI FABRIZIA ET AL: "DNA-based selection and screening of peptide ligands.", NATURE BIOTECHNOLOGY, vol. 16, no. 11, November 1998 (1998-11-01), pages 1068 - 1074, XP001030798, ISSN: 1087-0156 *
FELICI, FRANCO ET AL: "Mimicking of discontinuous epitopes by phage-displayed peptides II. Selection of clones recognized by a protective monoclonal antibody against the Bordetella pertussis toxin from phage peptide libraries", GENE (1993), 128(1), 21-7, XP000652423 *
PREZZI C ET AL: "Selection of antigenic and immunogenic mimics of hepatitis C virus using sera from patients", JOURNAL OF IMMUNOLOGY, THE WILLIAMS AND WILKINS CO. BALTIMORE, US, vol. 156, 1996, pages 4504 - 4513, XP002111267, ISSN: 0022-1767 *
PUNTORIERO GIULIA ET AL: "Towards a solution for hepatitis C virus hypervariability: Mimotopes of the hypervariable region 1 can induce antibodies cross-reacting with a large number of viral variants.", EMBO (EUROPEAN MOLECULAR BIOLOGY ORGANIZATION) JOURNAL, vol. 17, no. 13, 1 July 1998 (1998-07-01), pages 3521 - 3533, XP002111268, ISSN: 0261-4189 *
SAGGIO, ISABELLA ET AL: "Biotin binders selected from a random peptide library expressed on phage", BIOCHEM. J. (1993), 293(3), 613-16, XP000652418 *
URBANELLI L ET AL: "''Affinity maturation'' of ligands for HCV-specific serum antibodies", JOURNAL OF IMMUNOLOGICAL METHODS, ELSEVIER SCIENCE PUBLISHERS B.V.,AMSTERDAM, NL, vol. 236, no. 1-2, March 2000 (2000-03-01), pages 167 - 176, XP004190755, ISSN: 0022-1759 *
ZHU ZONG YU ET AL: "'In vitro evolution' of ligands for HCV-specific serum antibodies.", BIOLOGICAL CHEMISTRY, vol. 381, no. 3, March 2000 (2000-03-01), pages 245 - 254, XP001035267, ISSN: 1431-6730 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2108656A1 (fr) 2008-03-19 2009-10-14 Beninati, Concetta Fragments de protéines antigéniques de streptococcus pneumoniae
WO2009158682A2 (fr) * 2008-06-27 2009-12-30 Watkinson D Tobin Compositions et procédés pour diagnostiquer et traiter des troubles pathogènes
WO2009158682A3 (fr) * 2008-06-27 2010-07-22 Watkinson D Tobin Compositions et procédés pour diagnostiquer et traiter des troubles pathogènes

Also Published As

Publication number Publication date
SK5362003A3 (en) 2003-09-11
PL364832A1 (en) 2004-12-27
EP1332369A1 (fr) 2003-08-06
CA2427602A1 (fr) 2002-05-10
KR20030084895A (ko) 2003-11-01
HUP0302103A3 (en) 2007-03-28
CN1455866A (zh) 2003-11-12
JP2004513346A (ja) 2004-04-30
AU2001218836A1 (en) 2002-05-15
BR0017366A (pt) 2004-06-15
MXPA03003794A (es) 2004-10-15
HUP0302103A2 (hu) 2003-09-29

Similar Documents

Publication Publication Date Title
Folgori et al. A general strategy to identify mimotopes of pathological antigens using only random peptide libraries and human sera.
Prezzi et al. Selection of antigenic and immunogenic mimics of hepatitis C virus using sera from patients.
JP3188717B2 (ja) Ｃ型肝炎アッセイ
JP3645904B2 (ja) Ｃ型肝炎ウイルスの型の分類方法およびそこで使用する試薬
US6291197B1 (en) Binding moieties for human parvovirus B19
US5229491A (en) Peptides immunochemically reactive with antibodies directed against hepatitis non-a, non-b virus
Liu et al. Disease-specific B Cell epitopes for serum antibodies from patients with severe acute respiratory syndrome (SARS) and serologic detection of SARS antibodies by epitope-based peptide antigens
JPH08510063A (ja) リコンビナントタンパクおよび合成ペプチド試薬を用いるｈｉｖイムノアッセイ
JPH10502165A (ja) 合成ペプチド−改良イムノアッセイを使用する異なるｈｉｖ遺伝子型の検出
WO2002037115A1 (fr) Détection d'agents infectieux au moyen de mimiques d'antigènes
Minenkova et al. ADAM‐HCV, a new‐concept diagnostic assay for antibodies to hepatitis C virus in serum
EP0525910A1 (fr) Peptide non-A, non-B
US5985541A (en) Peptide capable of being recognized by antibodies recognizing the C33 antigen of hepatitis C virus
JP3022954B2 (ja) タイプ特異的抗原を使用する血清学的タイプ分け方法
JPH06510988A (ja) 合成ペプチドを用いてのｈｔｌｖ−１とｈｔｌｖ−２との区別
JPH05331193A (ja) 非ａ非ｂ型肝炎ウイルスに対する抗体と免疫化学的に反応するペプチド
JPH05320192A (ja) 非ａ非ｂ型肝炎ウイルスに対する抗体に免疫化学反応性を示すペプチド
EP0536838A2 (fr) Peptides non-A, non-B
CZ20031044A3 (cs) Způsob diagnostikování infekčních agens s využitím antigenových mimetik
JPH05271277A (ja) 非−ａ、非−ｂ肝炎ウイルスに対する抗体と免疫化学的に反応するペプチド
JP2655205B2 (ja) 非ａ非ｂ肝炎のアッセイ
WO1993011158A2 (fr) Peptides non-a, non-b
JPH04221398A (ja) 非ａ非ｂ型肝炎ウイルスに対する抗体と免疫化学反応するペプチド
CA2162250C (fr) Methodes de typage du virus de l'hepatite c et reactifs utiles a cette fin
CN118725022A (zh) 可中和新型冠状病毒的多肽序列及筛选方法和应用

Legal Events

Date Code Title Description
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: PV2003-1044

Country of ref document: CZ

WWE Wipo information: entry into national phase

Ref document number: 2001218836

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: PA/a/2003/003794

Country of ref document: MX

Ref document number: 008200009

Country of ref document: CN

WWE Wipo information: entry into national phase

Ref document number: 2427602

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 5362003

Country of ref document: SK

Ref document number: 2002539818

Country of ref document: JP

Ref document number: 1020037006153

Country of ref document: KR

WWE Wipo information: entry into national phase

Ref document number: 2000981607

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 2000981607

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWP Wipo information: published in national office

Ref document number: PV2003-1044

Country of ref document: CZ

WWP Wipo information: published in national office

Ref document number: 1020037006153

Country of ref document: KR

WWW Wipo information: withdrawn in national office

Ref document number: 2000981607

Country of ref document: EP