CN108727496A - The anti-human p63 protein monoclonal antibodies of mouse prepare and its immunohistochemistry purposes - Google Patents
The anti-human p63 protein monoclonal antibodies of mouse prepare and its immunohistochemistry purposes Download PDFInfo
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Abstract
The invention discloses a kind of monoclonal antibody of the anti-human p63 albumen of mouse, specificity is high, performance is stablized and potency is high, can be used for detecting the expression of p63 albumen in epithelial malignancy, and there is certain medical basic research to be worth.The invention further relates to application of the said monoclonal antibody in preparing the immunohistochemistry detection instrument for detecting p63 albumen.The present invention establishes the immunohistochemistry technology based on p63 monoclonal antibodies, for the auxiliary diagnosis of epithelial malignancy, therapeutic scheme selection and prognosis, epithelial malignancy includes mainly squamous cell carcinoma, cervical carcinoma, cervical squamous cell carcinoma, lung squamous cancer, adenocarcinoma of endometrium, gastric cancer, prostate cancer, non-small cell lung cancer, non-Hodgkin lymphoma etc..
Description
Technical field
The present invention relates to field of immunology, more particularly to the monoclonal antibody and the antibody of a kind of anti-human p63 albumen of mouse
Immunohistochemistry purposes.
Background technology
P63 is one of p53 family members, and family member further includes p73 and p53, although p63 is found later,
Evolutionary analysis shows that p73 and p53 is the evolution product of p63.P63 contains 3 conserved domains：Transcription activating domain（TA）,DNA
Binding domain（DBD）With oligomerization binding domain（OD）.P63 has similar signal path in structure with p53 very high homologies, the two
The functions such as inducing cell apoptosis, arresting cell cycle, while respectively possessing entirely different physiologic function again.People source p63 bases
Because being located at chromosome 3q27-29, including 15 exons, and contain 2 promoters.According to promoter（Transcripting start point）
Difference, codified generates two kinds of isomers with different activities：A kind of subtype protein TAp63 for full-length（N-terminal has
There is TA）, by the transcription of trans-activation p53 target genes, inducing cell apoptosis and aging finally play p53 sample cancer suppressing actions；
Another kind is the subtype protein Δ Np63 for the Truncated that N-terminal lacks TA, can inhibit the transcriptional activity of TAp63 and p53, promotes cell
Proliferation, tumour occur.Research shows that two kinds of isomers all have the effect for inhibiting Malignant tumor of bonal metastasis, in tissue extensively
Expression, and be selectively expressed in lung, skin, bone, muscle, mammary gland, thymus gland, lymphocyte, nerve fiber, digestive system and
Urogenital system etc., it is especially common in the epithelial cell of hyperplasia.Therefore, p63 gives birth to as the molecule of epithelial tissue stem cell
Object marks, and it is pre- for the pathogenesis of malignant tumour and treatment to study its expression in the malignant tumour in epithelial tissue source
There is important scientific meaning and clinical value afterwards.
In recent years, p63 is carried out large sample analysis by virologist in epithelial tumor tissue, it is found that p63 is on uterine neck
One of infected mark of chrotoplast, illustrates that it plays an important role during the occurrence and development of cervical carcinoma, p63 is prompted to be expected to
As the biological marker of cervical carcinoma and precancerous lesion.Some scholars find that p63 may be a valuable diagnosis lung squamous cancer
With the marker for judging squamous carcinoma differentiation degree.Currently, the p63 antibody researched and developed both at home and abroad is less, in terms of specificity and affinity
It is to be improved, therefore the urgently new anti-p63 monoclonal antibodies haveing excellent performance of research and development, the blank of the domestic industry is filled up, is reduced
Medical expense mitigates patient's burden.The present invention develops a high species specificity, performance stabilization and the high anti-p63 albumen of potency
Monoclonal antibody specific, can be used for malignant tumour occur, progress and prognosis in the relevant pathological studies of p63, be clinical treatment
New approaches are provided, there is certain medical basic research to be worth.The present invention uses immunohistochemistry（IHC）Method is established and is based on institute
State the immunohistochemistry technology of p63 monoclonal antibodies, for epithelial malignancy auxiliary diagnosis, therapeutic scheme selection and
Prognosis, malignant tumour include mainly squamous cell carcinoma, cervical carcinoma, cervical squamous cell carcinoma, lung squamous cancer, adenocarcinoma of endometrium, gastric cancer, forefront
Gland cancer, non-small cell lung cancer, non-Hodgkin lymphoma etc..
Invention content
Based on above-described state of the art, the purpose of the present invention be provide a species specificity it is good, performance stablize and
The monoclonal antibody of the high anti-p63 albumen of potency, and establish the immunohistochemistry technology based on the p63 monoclonal antibodies.
To achieve the above object, the present invention adopts the following technical scheme that：
The preparation method of p63 monoclonal antibodies of the present invention is as follows：
（1）The structure of recombinant expression carrier：According to the ORF complete sequences of people's p63 genes（Uniprot-Q9H3D4）, order gene and close
At acquisition p63 genetic immunizations source target sequence（P63 nucleotide sequences and amino acid sequence are shown in sequence table respectively）, gene both sides point
Not Yin Ru restriction endonuclease sites NheI and XhoI, be inserted into expression vector pDB-His-GST and pDB-His-MBP（Canada
Using biomaterial Co., Ltd, Applied Biological Materials Inc.）, build p63 recombinant expression plasmids
（P63-pDB-GST and p63-pDB-MBP）.The former p63-pDB-GST uses to immunization experiment animal, the latter p63-pDB-MBP
With ELISA screening positive clones.
（2）The expression and purification of p63 recombinant proteins：By p63-pDB-GST recombinant expression plasmids conversion BL21 DE3 Star
Competent cell, centrifuging and taking supernatant after lytic cell, Nickel column purifications obtain the p63 recombinant proteins of purifying.
（3）The screening and preparation of monoclonal antibody：BALB/c mouse are immunized using the p63-pDB-GST recombinant proteins of above-mentioned purifying,
Mouse spleen cells are taken to be merged with SP2/0 cells, limiting dilution assay obtains monoclonal, is overlay using p63-pDB-MBP
96 orifice plates screen positive hybridoma cell using ELISA methods, obtain the hybridoma cell strain that can secrete anti-p63 specific antibodies；
Mouse ascites are prepared, p63 monoclonal antibodies are purified by protein A column chromatographies.Pass through Western Blot respectively（The result is shown in Figure 1）
And immunohistochemical experiment（As a result see Fig. 2）Verify specificity and the sensitivity of the monoclonal antibody.
Meanwhile the present invention provides a kind of said monoclonal antibody and is preparing the immunohistochemistry tool for detecting p63 albumen
In application.
The immune detection tool is reagent, kit, chip or test strips.
The present invention also provides said monoclonal antibodies to prepare the immunohistochemistry use for detecting epithelial malignancy
On the way.Mark of molecular biology of the p63 as epithelial tissue stem cell, research prompt may be a valuable diagnosing cervical
With the biological marker of lung squamous cancer.Therefore, expression of the p63 in tumour cell is detected, for the auxiliary of body epithelial malignancy
Help diagnosis, therapeutic scheme selection and prognosis that there is important directive function.
By taking the immunohistochemistry automatic staining machine BondMax for using Leica as an example, exempted from using the p63 monoclonal antibodies
The condition of epidemic disease histochemical staining is：
（1）The IHC protocol F carried using machine are shown peroxidase closing step by using primary antibody to move forward to DAB
Before color；
（2）Antibody uses final concentration of 0.5 μ g/ml, is incubated at room temperature 30min；
（3）Antigen retrieval uses the antigen retrieval buffers of pH 9.0（ER2）, 100 DEG C of incubation with heat 30min.
When using other IHC automatic staining machines or carrying out hand dyeing, above-mentioned condition progress is please referred to.
Specific monoclonal antibody of the present invention while can be established based on the monoclonal antibody with p63 albumen specific bonds
Immunohistochemistry technology, can be used for epithelial malignancy auxiliary diagnosis, therapeutic scheme selection and prognosis, malignant tumour
Include mainly squamous cell carcinoma, cervical carcinoma, cervical squamous cell carcinoma, lung squamous cancer, adenocarcinoma of endometrium, gastric cancer, prostate cancer, non-small cell
Lung cancer, non-Hodgkin lymphoma etc..
Description of the drawings
Fig. 1 is the Western blot results of monoclonal antibody of the present invention：M swimming lanes are Marker, and 1 swimming lane is p63 genes
The 293T HEK cell pyrolysis liquids of transfection, 2 swimming lanes are the CHO-K1 cell pyrolysis liquids of p63 genes transfection, and 3 swimming lanes are untransfected
Negative control 293T HEK cell pyrolysis liquids.Wherein A figures be using p63 monoclonal antibodies of the present invention detect as a result, B figures are to make
With the result of the positive control of β-Actin antibody tests.
Fig. 2 is using monoclonal antibody of the present invention as primary antibody, and Immunohistochemical Method detects p63 albumen in human thyroid carcinoma
Express colored graph.
Specific implementation mode
The present invention disclose a species specificity it is good, performance stablize and the high anti-p63 albumen of potency monoclonal antibody, Yi Jijian
Survey the immunohistochemistry purposes of tumour cell p63 albumen.Those skilled in the art can refer to present disclosure and are suitably modified with reality
Now and using technology of the invention, but similar replacement and change should be regarded as being included in the present invention.
In order to make those skilled in the art more fully understand technical scheme of the present invention, with reference to specific embodiment pair
The present invention is described in further detail.
The preparation of 1 anti-p63 monoclonal antibodies of embodiment
One, the structure of p63 recombinant expression plasmids
The p63 genes reported according to uniprot（Uniprot-Q9H3D4）, order gene chemical synthesis and obtain p63 genetic immunizations source mesh
Sequence is marked, and restriction enzyme site NheI and XhoI are introduced respectively at sequence both ends, and then is cloned into expression vector pDB-His- respectively
GST and pDB-His-MBP plasmids establish p63-pDB-GST and p63-pDB-MBP recombinant expression plasmids, are respectively used to immune real
Test animal and ELISA screening positive clones.
Two, the expression and purification of p63 recombinant proteins
1, BL21 DE3 Star competent cells are converted
It is thin that it is transformed into BL21 DE3 Star competence respectively using p63-pDB-GST the and p63-pDB-MBP plasmids of above-mentioned structure
Born of the same parents, preculture obtain 250ml bacterium liquid, and 6L further, which is added, in pre-culture solution contains kanamycins（Kanamycin）LB culture
In liquid, 37 DEG C of cultures to OD values, using the IPTG induced expressions of 0.5mM, cultivate 4h up to 0.8.
2, lytic cell
After centrifugation obtains cell precipitation, with solution A（20mM sodium phosphates, pH 8.0,500mM sodium chloride solutions）Cell is resuspended,
Ultrasonication 1min x 6 times.3000 g/min centrifuge 15min sedimentation cell fragments, collect supernatant for purifying in next step.
3, Nickel column chromatographies purify
With aperture for 0.45 μm, lysate supernatant after the pvdf membrane filter filter centrifugation of a diameter of 33mm is simultaneously transferred to 15ml pipes.
10 ml solution As are added and are mixed with sample and prepared column purification.After Nickel resins overturn mixing repeatedly, 3ml is taken to be added in column,
It is cleaned and is balanced 3 times with solution A, standing makes resin natural sediment.Sample is added, collects efflux for the first time.Column is cleaned with solution A
5 times of preconfigured eluents of volume are added in son 5 times（250mM imidazoles is added in solution A）.Eluent is collected after standing 1min.
4. dialysis
It is dialysed using 1 × PBS solution by the sample that step 3 elutes, make two bites at a cherry dialysis, shares 4L dialyzates.
Three, the screening and preparation of monoclonal antibody
1, animal immune：Above-mentioned purified p63-pDB-GST recombinant proteins are emulsified with complete Freund's adjuvant, using subcutaneous or
6-8 week old BALB/c mouses are immunized in intraperitoneal injection method, and only for 50 μ g/, interval carries out being immunized for second after two weeks immunizing dose,
It is emulsified with incomplete Freund's adjuvant, immunizing dose is 50 μ g/.It is immune that tail blood is taken to be measured with ELISA method gradient dilution afterwards twice
Serum titer；Determine whether booster immunization according to result, chooses the highest mouse of antibody titer and carry out cell fusion.
2, cell fusion：Myeloma cell uses the SP2/0 in the sources BALB/c, and exponential phase is in when fusion；It takes
Immune mouse spleen is stated, lymphocyte single cell suspension is made；Immune mouse spleen lymphocyte is with myeloma cell with 1:5-
1:10 mixing, are added dropwise 37 DEG C of 50% PEG（pH 8.0）1ml, is added incomplete culture medium and remaining terminate liquid, and centrifugation is abandoned
HAT culture medium suspension mixings are added after clear, MC constant volumes to 50ml are dispensed into 3.5cm culture dishes, are put in wet box, are placed in 37
DEG C, 5% CO2It is cultivated in constant incubator.
3, it screens and clones：Fusion selects cell clone in 7-10 days, and egg is recombinated using the p63-pDB-MBP of above-mentioned purifying
It is white to carry out ELISA tests, mark cell strain number.Limiting dilution is carried out to positive hole cell, is measured within 5-6 days after each limiting dilution
ELISA values, the picking OD280 positives are worth higher monoclonal hole and carry out limiting dilution, until ELISA measures 96 orifice plates hardened fruit entirely
For the positive.
4, the preparation and purification of ascites monoclonal antibodies：0.5ml norphytanes are injected intraperitoneally in the male BALB/c mouse of 10-12 week old,
Every mouse is injected intraperitoneally with 1ml syringes and wash the monoclonal cell suspension being resuspended through PBS after a week, cell dosage for 5 ×
106/ only, make a call to 2 mouse per strain antibody.Ascites, centrifuging and taking supernatant, protein A column chromatographies are collected after mouse ascites accumulation
Method carries out ascites purifying, and monoclonal antibody concentration mensuration after purification is dispensed, frozen at -20 DEG C.
Using monoclonal antibody of the present invention as primary antibody, to the 293T HEK cells and CHO-K1 cells by the transfection of p63 genes
The protein lysate of system is hybridized, is developed the color, and the results are shown in Figure 1, as seen from the figure：The antibody successfully detects p63 genes, and
Single band is presented.
Embodiment 2 is using monoclonal antibody of the present invention as the immunohistochemical experiment of primary antibody
1, it takes 24 kinds of different types of cancerous tissue samplings to make organization chip respectively, uses Leica RM2235 type histotomes
It is sliced, slice thickness is 4 μm；
2, immunohistochemical staining test is carried out to antibody of the present invention using Leica BondMax immunohistochemistry automatic staining machines, made
The dewaxing carried with machine and hydrating condition, the specific steps are：60 DEG C of incubation 30min, using dewaxed solution（Leica）It washes 3 times.
Antigen retrieval uses antigen retrieval buffers 2（ER2, Leica）, 100 DEG C of incubation 30min.Primary antibody uses antibody of the present invention, using antibody
Dilution（Leica）It is diluted to final concentration of 0.5 μ g/ml, 150 μ l.Antibody at room temperature is incubated 30min.Use mating secondary antibody
（Leica）150 μ l are incubated at room temperature 8min.Use poly analyte detection liquid（Leica）150 μ l are incubated at room temperature 8min.Using endogenous
Property peroxidase confining liquid 150 μ l be incubated at room temperature 5min.Use DAB developing solutions（Leica）150 μ l are incubated at room temperature 10min.
Haematoxylin is redyed, and 5min is incubated at room temperature；
3, it is dehydrated and transparent：Deionized water cleans 1min, 95% ethyl alcohol 1min, 100% ethyl alcohol 2min x 2 times, dimethylbenzene 2min
X 3 times, neutral gum mounting；
4, microscopy, as a result：Partial tumors and normal epithelial tissues show nucleus positive dyeing, including cervical carcinoma etc., part sample
This display is negative to be dyed without p63.Wherein thyroid cancer dyeing is as shown in Figure 2, it is seen that apparent nuclear targeting.Staining pattern is just
Really, signal is stronger, has no apparent non-specific dyeing.
The specific detection of 3 monoclonal antibody of the present invention of embodiment
96 orifice plates overlay using ELISA detection irrelevant antigens using antibody of the present invention（Her-2）, result is feminine gender.
The above is only a preferred embodiment of the present invention, it should be appreciated that for those of ordinary skill in the art,
Various improvements and modifications may be made without departing from the principle of the present invention, and such equivalent forms also should be regarded as this hair
Bright protection domain.
<110>Nanjing Jino rice medical science and technology Co., Ltd
<120>The anti-human p63 protein monoclonal antibodies of mouse prepare and its immunohistochemistry purposes
<210> 1
<211>2043
<212> DNA
<213>Artificial sequence
<400> 1
ORIGIN
1 ATGAATTTTG AAACTTCACG GTGTGCCACC CTACAGTACT GCCCTGACCC TTACATCCAG
61 CGTTTCGTAG AAACCCCAGC TCATTTCTCT TGGAAAGAAA GTTATTACCG ATCCACCATG
121 TCCCAGAGCA CACAGACAAA TGAATTCCTC AGTCCAGAGG TTTTCCAGCA TATCTGGGAT
181 TTTCTGGAAC AGCCTATATG TTCAGTTCAG CCCATTGACT TGAACTTTGT GGATGAACCA
241 TCAGAAGATG GTGCGACAAA CAAGATTGAG ATTAGCATGG ACTGTATCCG CATGCAGGAC
301 TCGGACCTGA GTGACCCCAT GTGGCCACAG TACACGAACC TGGGGCTCCT GAACAGCATG
361 GACCAGCAGA TTCAGAACGG CTCCTCGTCC ACCAGTCCCT ATAACACAGA CCACGCGCAG
421 AACAGCGTCA CGGCGCCCTC GCCCTACGCA CAGCCCAGCT CCACCTTCGA TGCTCTCTCT
481 CCATCACCCG CCATCCCCTC CAACACCGAC TACCCAGGCC CGCACAGTTT CGACGTGTCC
541 TTCCAGCAGT CGAGCACCGC CAAGTCGGCC ACCTGGACGT ATTCCACTGA ACTGAAGAAA
601 CTCTACTGCC AAATTGCAAA GACATGCCCC ATCCAGATCA AGGTGATGAC CCCACCTCCT
661 CAGGGAGCTG TTATCCGCGC CATGCCTGTC TACAAAAAAG CTGAGCACGT CACGGAGGTG
721 GTGAAGCGGT GCCCCAACCA TGAGCTGAGC CGTGAATTCA ACGAGGGACA GATTGCCCCT
781 CCTAGTCATT TGATTCGAGT AGAGGGGAAC AGCCATGCCC AGTATGTAGA AGATCCCATC
841 ACAGGAAGAC AGAGTGTGCT GGTACCTTAT GAGCCACCCC AGGTTGGCAC TGAATTCACG
901 ACAGTCTTGT ACAATTTCAT GTGTAACAGC AGTTGTGTTG GAGGGATGAA CCGCCGTCCA
961 ATTTTAATCA TTGTTACTCT GGAAACCAGA GATGGGCAAG TCCTGGGCCG ACGCTGCTTT
1021 GAGGCCCGGA TCTGTGCTTG CCCAGGAAGA GACAGGAAGG CGGATGAAGA TAGCATCAGA
1081 AAGCAGCAAG TTTCGGACAG TACAAAGAAC GGTGATGGTA CGAAGCGCCC GTTTCGTCAG
1141 AACACACATG GTATCCAGAT GACATCCATC AAGAAACGAA GATCCCCAGA TGATGAACTG
1201 TTATACTTAC CAGTGAGGGG CCGTGAGACT TATGAAATGC TGTTGAAGAT CAAAGAGTCC
1261 CTGGAACTCA TGCAGTACCT TCCTCAGCAC ACAATTGAAA CGTACAGGCA ACAGCAACAG
1321 CAGCAGCACC AGCACTTACT TCAGAAACAG ACCTCAATAC AGTCTCCATC TTCATATGGT
1381 AACAGCTCCC CACCTCTGAA CAAAATGAAC AGCATGAACA AGCTGCCTTC TGTGAGCCAG
1441 CTTATCAACC CTCAGCAGCG CAACGCCCTC ACTCCTACAA CCATTCCTGA TGGCATGGGA
1501 GCCAACATTC CCATGATGGG CACCCACATG CCAATGGCTG GAGACATGAA TGGACTCAGC
1561 CCCACCCAGG CACTCCCTCC CCCACTCTCC ATGCCATCCA CCTCCCACTG CACACCCCCA
1621 CCTCCGTATC CCACAGATTG CAGCATTGTC AGTTTCTTAG CGAGGTTGGG CTGTTCATCA
1681 TGTCTGGACT ATTTCACGAC CCAGGGGCTG ACCACCATCT ATCAGATTGA GCATTACTCC
1741 ATGGATGATC TGGCAAGTCT GAAAATCCCT GAGCAATTTC GACATGCGAT CTGGAAGGGC
1801 ATCCTGGAC CACCGGCAGCT CCACGAATTC TCCTCCCCTT CTCATCTCCT GCGGACCCCA
1861 AGCAGTGCC TCTACAGTCAG TGTGGGCTCC AGTGAGACCC GGGGTGAGCG TGTTATTGAT
1921 GCTGTGCGA TTCACCCTCCG CCAGACCATC TCTTTCCCAC CCCGAGATGA GTGGAATGAC
1981 TTCAACTTT GACATGGATGC TCGCCGCAAT AAGCAACAGC GCATCAAAGA GGAGGGGGAG
2041 TGA
//
<210> 2
<211>680
<212> PRT
<213>Artificial sequence
<400> 2
ORIGIN
1 MNFETSRCAT LQYCPDPYIQ RFVETPAHFS WKESYYRSTM SQSTQTNEFL SPEVFQHIWD
61 FLEQPICSVQ PIDLNFVDEP SEDGATNKIE ISMDCIRMQD SDLSDPMWPQ YTNLGLLNSM
121 DQQIQNGSSS TSPYNTDHAQ NSVTAPSPYA QPSSTFDALS PSPAIPSNTD YPGPHSFDVS
181 FQQSSTAKSA TWTYSTELKK LYCQIAKTCP IQIKVMTPPP QGAVIRAMPV YKKAEHVTEV
241 VKRCPNHELS REFNEGQIAP PSHLIRVEGN SHAQYVEDPI TGRQSVLVPY EPPQVGTEFT
301 TVLYNFMCNS SCVGGMNRRP ILIIVTLETR DGQVLGRRCF EARICACPGR DRKADEDSIR
361 KQQVSDSTKN GDGTKRPFRQ NTHGIQMTSI KKRRSPDDEL LYLPVRGRET YEMLLKIKES
421 LELMQYLPQH TIETYRQQQQ QQHQHLLQKQ TSIQSPSSYG NSSPPLNKMN SMNKLPSVSQ
481 LINPQQRNAL TPTTIPDGMG ANIPMMGTHM PMAGDMNGLS PTQALPPPLS MPSTSHCTPP
541 PPYPTDCSIV SFLARLGCSS CLDYFTTQGL TTIYQIEHYS MDDLASLKIP EQFRHAIWKG
601 ILDHRQLHEF SSPSHLLRTP SSASTVSVGS SETRGERVID AVRFTLRQTI SFPPRDEWND
661 FNFDMDARRN KQQRIKEEGE
//
Claims (6)
1. a kind of mouse anti-human monoclonal's antibody, it is characterised in that people's p63 albumen can be specifically bound.
2. monoclonal antibody described in claim 1 is preparing the immunohistochemistry detection instrument for specific detection p63 albumen
In application.
3. application according to claim 2, which is characterized in that the immunohistochemistry detection instrument is reagent, kit, core
Piece or test strips.
4. application according to claim 2, which is characterized in that the immunohistochemistry detection instrument can be used for basic medical and grind
Study carefully and there is important clinical reference value to the diagnosis of epithelial malignancy, the selection of therapeutic scheme and prognosis.
5. application according to claim 4, which is characterized in that the medical basic research is that p63 albumen swells with pernicious
Tumor generation, progress and the relevant pathological study of prognosis.
6. application according to claim 4, which is characterized in that the epithelial malignancy includes mainly pinacocyte
Cancer, cervical carcinoma, cervical squamous cell carcinoma, lung squamous cancer, adenocarcinoma of endometrium, gastric cancer, prostate cancer, non-small cell lung cancer, non-Hodgkin's leaching
Bar tumor etc..
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102924594A (en) * | 2012-08-03 | 2013-02-13 | 无锡傲锐东源生物科技有限公司 | Anti-tumor-protein-P63 monoclonal antibody and purpose thereof |
| CN104447994A (en) * | 2014-12-05 | 2015-03-25 | 福州迈新生物技术开发有限公司 | Tumor suppressor gene p63 monoclonal antibody and application thereof |
| EP2900265A1 (en) * | 2012-09-27 | 2015-08-05 | Biocare Medical, LLC | Anti-uroplakin ii antibodies systems and methods |
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2018
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102924594A (en) * | 2012-08-03 | 2013-02-13 | 无锡傲锐东源生物科技有限公司 | Anti-tumor-protein-P63 monoclonal antibody and purpose thereof |
| EP2900265A1 (en) * | 2012-09-27 | 2015-08-05 | Biocare Medical, LLC | Anti-uroplakin ii antibodies systems and methods |
| CN104447994A (en) * | 2014-12-05 | 2015-03-25 | 福州迈新生物技术开发有限公司 | Tumor suppressor gene p63 monoclonal antibody and application thereof |
Non-Patent Citations (1)
| Title |
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