CN101775371B - Anti-testicle-protein hTSC29 antibody or antibody fragment and application thereof - Google Patents
Anti-testicle-protein hTSC29 antibody or antibody fragment and application thereof Download PDFInfo
- Publication number
- CN101775371B CN101775371B CN2009101886409A CN200910188640A CN101775371B CN 101775371 B CN101775371 B CN 101775371B CN 2009101886409 A CN2009101886409 A CN 2009101886409A CN 200910188640 A CN200910188640 A CN 200910188640A CN 101775371 B CN101775371 B CN 101775371B
- Authority
- CN
- China
- Prior art keywords
- antibody
- htsc29
- protein
- testicle
- mab
- Prior art date
- Legal status (The legal status 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 status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses an anti-testicle-protein hTSC29 antibody or an antibody fragment and application thereof. The anti-testicle-protein hTSC29 antibody or the antibody fragment is secreted by a hybridoma strain hTSC29 with the preserving number of CCTCC NO: C200952. Through the antibody or the antibody fragment, people can know molecular mechanisms formed by human sperms, and can provide the specificity detection for parts of male infertility.
Description
Technical field
The invention relates to a kind of antibody or antibody fragment and application thereof that male fertility detects that be used for.
Background technology
In worldwide, about 15% Mr. and Mrs receive the influence of infertility, and wherein male fertility has become a main healthy reproduction problem.Along with the raising of social industrialization degree and the quickening of rhythm of life; Add factors such as environmental pollution and bad life habits; The semen quality descends day by day, and the male sterility number is ascendant trend year by year, and the problem of male reproductive health more and more receives the concern of the whole society.Spermatogeny is the process of a complicacy, and it is divided into spermatogonium mitotic division, spermatocyte reduction division and sperm and forms three phases.This unique biological phenomena in the body needs the participation of a series of testes specificity expressing genes, is bringing into play particular functionality separately.More known genes and spermatogeny are closely related at present; CREM for example, Ddrl, TESP-1 etc. all are proved to be for the testes specificity expressing gene and in the spermatogeny process and play an important role; The disappearance of these genes or sudden change will cause serious oligospermatism or azoospermia, thereby cause sterile.Therefore, screening and evaluation production of sperm genes involved help further to understand the molecular mechanism of human sperm's formation and the reason of part male sterility.
Summary of the invention
Technical problem to be solved by this invention is that a kind of antibody or antibody fragment and application thereof that can be used in the anti-testicle-protein hTSC 29 of male fertility detection is provided.
For solving the problems of the technologies described above, the present invention provides a kind of isolating hybridoma cell strain, and its preserving number is CCTCC NO:C200952.
A kind of by hybridoma cell strain hTSC29 excretory antibody or antibody fragment.
Further, antibody or antibody fragment are antibody or the antibody fragment that specificity combines anti-testicle-protein hTSC 29.This hybridoma cell strain hTSC29 also can secrete and can specificity combine other proteic antibody or antibody fragment.
Further, antibody is that specificity combines proteic polyclonal antibody of hTSC29 or monoclonal antibody.
Further, monoclonal antibody is IgG, and subclass is IgG2b (κ).
A kind of antibody of anti-testicle-protein hTSC 29 or the application of antibody fragment, said antibody or antibody fragment and coupling matter coupling join, and said coupling matter is enzyme, fluorescent marker, chemiluminescent labels or medicine.
A kind of antibody of anti-testicle-protein hTSC 29 or antibody fragment complex comprise that the antibody of anti-testicle-protein hTSC 29 or antibody reach the coupling matter that joins with its coupling, and said coupling matter is selected from affinity tag, medicine.
Said affinity tag is enzyme, fluorescent marker or chemiluminescent labels.
A kind of pharmaceutical composition comprises pharmaceutically antibody or the antibody fragment and the pharmaceutically acceptable carrier of the anti-testicle-protein hTSC 29 of significant quantity.
The application on male sterility detects of a kind of antibody of anti-testicle-protein hTSC 29 or antibody fragment.
The invention has the beneficial effects as follows that the applicant filters out people's testicle specificity gene hTSC29, take all factors into consideration, choose the 224-241 amino acids sequence of hTSC29 at last and synthesize according to the result of parameter predictions such as homology, antigenicity, wetting ability.Through mouse immune, cytogamy, screening and cloning, obtained the hybridoma cell strain of the anti-hTSC29 mAb of ability stably excreting, Western blot identifies and shows that this mAb is relative molecular mass (M in people's normal testis albumen
r) be about the 60kDa place and detect specific band; Immunohistochemistry shows that hTSC29 albumen mainly expresses in the spermatocyte of normal people's testis and round spermatid; This antibody titer is high, high specificity, for further studying the male fertility relevant with hTSC29 genetic expression special testing tool is provided.
Description of drawings
Fig. 1 is the specificity figure that Western blot detects hTSC29 mAb, wherein, character 1 representative's endometrial cell, 2 represent the leukemia cell, 3 representative's testis tissues, GAPDH is as confidential reference items;
Fig. 2 is the immunohistochemical methods location map of hTSC29, and the arrow indication is a hTSC29 protein, Showed by immune group result, and the molecule that hTSC29 mAb is discerned mainly is positioned at the spermatocyte and the spermatid of normal people's testis tissue, the negative contrast of A;
Fig. 3 is the expression figure of immunofluorescence dyeing technology for detection hTSC29 albumen in testis tissue, wherein, the negative contrast of A, B is the location (arrow indication) of hTSC29 albumen in people's testis tissue.
Embodiment
The application's applicant adopts the adult testis genomic expression spectrum chip of Affyymetrix company; Successfully filter out a new gene of human testicle specificity; And with its called after hTSC29 (GenBank number of landing: BC032957); For this expression of gene characteristic of more deep research and the biological function in the spermatogeny process thereof, the applicant has prepared the monoclonal antibody of the synthetic peptide of mouse-anti hTSC29; The applicant carries out homology analysis to hTSC29 full-length proteins sequence and other testes specificity expressed proteins; Choose the low fragment of homology; With DNAstar software proteinic epitope and wetting ability are analyzed again; Result according to parameter predictions such as homology, antigenicity, wetting abilities takes all factors into consideration, and chooses the 224-241 amino acids sequence of hTSC29 at last and synthesizes.Through mouse immune, cytogamy, screening and cloning, obtained the hybridoma cell strain hTSC29 of the anti-hTSC29 mAb of ability stably excreting, the preserving number of this cell strain is CCTCC NO:C200952, and excretory antibody is IgG2b (κ) hypotype, and tiring reaches 1: 10
4Western blot identifies and shows that this mAb is relative molecular mass (M in people's normal testis albumen
r) be about the 60kDa place and detect specific band; Immunohistochemistry shows that hTSC29 albumen mainly expresses in the spermatocyte of normal people's testis and round spermatid; This antibody titer is high, high specificity, for further studying the male fertility relevant with hTSC29 genetic expression special testing tool is provided.
The present invention adopts bioinformatics method the protein sequence of hTSC29 is analyzed and to be predicted; Design one section polypeptide of forming by 19 amino acid (peptide sequence is SPENTCPREATKKSRHGLD) according to physico-chemical properties such as homology, antigenicity, wetting abilities; With itself and carrier proteins keyhole limpet hemocyanin (keyhole limpet hemocyanin; KLH) coupling pneumoretroperitoneum injecting immune BALB/c mouse; Use the anti-hTSC29 monoclonal antibody of hybridoma technology preparation; Use this new antibodies, through proteinic existence of method human body hTSC29 and expression characteristics thereof such as ELISA method, Western blot, immunohistochemistry and immunofluorescence dyeings.
The preservation information of hybridoma cell strain hTSC29 is following:
Culture title: hybridoma cell strain hTSC29
Preservation date: on August 27th, 2009
Depositary institution: Chinese typical culture collection center (CCTCC)
Deposit number: CCTCC NO:C200952
The preparation method of proteic antibody of anti-hTSC29 or antibody fragment is following:
1. material
Sp2/0 is available from ATCC in the murine myeloma cell strain; High sugared DMEM and foetal calf serum are available from Gibco; Freund adjuvant, incomplete freund adjuvant, HAT, HT and PEG (1450) are available from U.S. Sigma company fully; The HRP-goat anti-mouse igg is available from KPL company; The sheep anti mouse two of FITC mark is anti-available from Sigma company; 3,3,5,5-TMB (TMB) is available from Ke Hua biotech firm; Bovine serum albumin (BSA) is available from Merck company; Mouse subclass (type) detection kit is available from HBT company; ECL luminescence reagent box is available from PIERCE company; Nitrocellulose filter (PVDF) is available from Millipore company; Other reagent are homemade analytical pure.Electrophoresis chamber, electrophoresis apparatus (Mini-protean Teta cell type), ELIASA (680 type) and fluoroscopic image scanner are the BIO-RAD product; Half-dried membrane-transferring device is a Wealtec corp product; BALB/c mouse (7～8 ages in week, female, cleaning level) is provided by Zhongshan University's Experimental Animal Center; Normal people's testis tissue is provided by BeiJing University ShenZhen Hospital Pathology Deparment; Polypeptide synthetic with and accomplish by Xi'an China occasion bio tech ltd with the coupling of KLH, BSA.
2.hTSC29 antigenic obtaining and animal immune
Utilize the DNAstar software package hTSC29 aminoacid sequence of different genera to be carried out hydrophilic and hydrophobic, antigenicity and the surface exposure property prediction of multisequencing comparison and hTSC29; One section aminoacid sequence choosing in the hTSC29 albumen entrusts Xi'an China occasion bio tech ltd synthetic, shows synthetic polypeptide fragment and theoretical M through mass spectroscopy
rUnanimity, reversed-phase HPLC detect purity more than 95%; The crosslinked employing LUTARALDEHYDE connection method of polypeptide and KLH and BSA adds the synthetic polypeptide of 5mg respectively among 7mg KLH and the BSA, slowly adds the glutaraldehyde solution (1mL) of freshly prepared 3g/L while vibrating, room temperature effect 2h.With pH8.5 borate buffer dialysed overnight.Be cross-linked to form polypeptide-K LH, polypeptide-BSA conjugate.Get 50 μ g polypeptide-K LH conjugates add 200 μ L sterilization PBS mix with isopyknic complete freund adjuvant carry out fully emulsified, the immune BALB/c mouse of abdominal injection.Every at a distance from 2 all booster immunizations 1 time after the first immunisation, dosage is the same, after the full freund adjuvant that toos many or too much for use is fully emulsified, injects mouse peritoneal.Blood is got in docking behind the 3rd booster immunization, and separation of serum is measured serum titer.Select the high mouse of serum titer,, get mouse boosting cell after 3 days and merge with the polypeptide-K LH that does not add adjuvant booster immunization 1 time again.
3. the foundation of hybridoma cell strain
(1) cytogamy: under the aseptic condition, the Sp2/0 cell of the growth of taking the logarithm, the centrifugal 8min of 1000r/min abandons supernatant, with counting behind the incomplete substratum suspendible of the DMEM cell, gets required cell count.Preparation immune mouse spleen cell suspension; With Sp2/0 cell and splenocyte mixed by 1: 5, to merge 37 ℃ of water-bath 500mL/L PEG effect under, fusion is the centrifugal 15min of 1200r/min afterwards; Abandon supernatant; Select nutrient solution suspendible gently with HAT, be seeded in the 96 porocyte culture plates of having cultivated feeder cell in advance, put 37 ℃, 50mL/L CO
2Cultivate in the incubator.Change HT on the 10th day and select nutrient solution, changed the DMEM complete culture solution that contains the 100mL/L foetal calf serum on the 14th day.
(2) screening and cloning: grow to hole floorage 1/10 when above when hybridoma after 2～3 weeks; Polypeptide-BSA conjugate with 5mg/L encapsulates polystyrene micropore plate, adds the hybridoma supernatant, 37 ℃ hatch 1h after; PBST (PBS+0.5mL/L Tween-20) washing; Add the HRP-goat anti-mouse igg antibody, after the TMB colour developing, measure A with ELIASA
450/630Value.With normal BALB/c mouse serum as negative control; Immune BALB/c mouse serum is as positive control; Positive with P/N＞2, adopt limiting dilution assay to clone 2～3 times continuously to choosing positive porocyte, reach 100% up to the mAb in clonal growth hole positive rate; Set up the monoclonal cell strain, this monoclonal cell strain is the cell strain that preserving number is CCTCC NO:C200952.After positive hybridoma cell strain enlarged culturing, carry out the preparation of cell cryopreservation and ascites.The monoclonal cell strain still keeps the ability of stably excreting mAb after frozen, recovery.
4.mAb the preparation of ascites and titration thereof
Every BALB/c mouse intraperitoneal injection 0.5mL sterilising liq paraffin, the hybridoma that will build strain after 7 days is with 1 * 10
6Individual cell inoculation is pressed conventional method and is collected, prepares mAb ascites in mouse peritoneal.Adopting above-mentioned ELISA method to detect mAb ascites tires.
5.mAb the IgG subgroup identification
Mouse mAb subclass detection kit specification sheets according to HBT company carries out.In the Glass tubing that is placed with inhomogeneity and subclass test strip respectively, add the detection antibody of damping fluid, mAb to be checked and respective class and subclass in succession, spot appears in tube shaken on test strip gently.The result shows hybridoma cell strain excretory mAb, and subclass is IgG2b (κ), and tiring reaches 1: 10
4
6.mAb specific detection
(1) Western blot analyzes: get normal people's testis tissue, be cut into fine debris, cell rinsing liquid rinsing 1 time; Added RIPA lysate and proteinase inhibitor 1: 100; The mixing tissue, the homogenate sample, hatch 30min on ice after; The centrifugal 20min of 12000g obtains normal people's testis tissue lysate.The density of treating endometrial cell and white blood disease K-562 cell reaches about 2 * 10
8During/L, obtain cell pyrolysis liquid with aforesaid method.Above-mentioned 3 kinds of albumen add 5 * SDS Loading buffer in proportion, 100 ℃ the heating 15min, carry out conventional SDS-PAGE electrophoresis after, semidrying goes on the pvdf membrane, spends the night with 4 ℃ of sealings of 10g/L BSA.Add the mAb of dilution in 1: 1000 next day, hatch 4h under the room temperature, TBST (TBS+0.5mL/L Tween-20) damping fluid is washed film 3 times; 10min/ time, add the sheep anti-mouse igg antibody of HRP mark of dilution in 1: 2000, hatch 1h under the room temperature; After TBST thoroughly washs; After the ECL agent treated, put on the fluoroscopic image scanner and make public, computingmachine will whenever be preserved at a distance from the detected signal of 20s clock automatically.
(2) immunohistochemical analysis: Paraformaldehyde 96 was fixed the routine paraffin wax embedded section after people's testis tissue was drawn materials.Section is through YLENE dewaxing, gradient alcohol aquation.Drip 30mL/L H
2O
2-methyl alcohol, the room temperature lucifuge is hatched 20min, to eliminate the catalatic effect of endogenous.The citrate buffer microwave heating of 10mmol/L (92 ℃～98 ℃, 15min) carry out antigen retrieval, seal 30min under the normal goats serum room temperature, drip the culture supernatant of hybridoma then successively, 4 ℃ of incubated overnight.It is anti-to add goat-anti mouse biotinylation two behind the PBS thorough washing, incubated at room 1h, and PBS washing 3 times adds the HRP-streptavidin, incubated at room 1h again.Carry out the DAB colour developing after PBS develops a film, Hematorylin is redyed, and blue back mounting is returned in the hydrochloride alcohol differentiation, and microscopically is observed, taken pictures.Negative control adopts normal mouse serum to replace the hybridoma supernatant.
(3) immunofluorescence analysis: normal people's testis tissue specimens paraffin embedding slices is with the conventional dewaxing of YLENE, entry, antigen retrieval.PBS washing 5 times drips 30mL/L H
2O
2-methyl alcohol, incubated at room 20min is to eliminate the catalatic effect of endogenous.PBS washing back drips under the normal goats serum room temperature seals 10min, drips the culture supernatant of hybridoma then successively, hatches 2h in the wet box.PBS washing 2 times, the sheep anti mouse two that adds the FITC mark is anti-, and lucifuge is hatched 30～40min.PBS washing 3 times, the glycerine mounting, fluorescence microscope is taken pictures.Negative control adopts normal mouse serum to replace the hybridoma supernatant.
7. antibody effect experiment
(1). the foundation of anti-hTSC29 mAb hybridoma cell strain
Through mouse immune, cytogamy, screening and cloning, obtain the positive colony of anti-hTSC29, carry out obtaining after 3 time cloningizations can stably excreting specific anti hTSC29 mAb hybridoma cell strain, its subclone is called after 4F9,2F7 and 4F2 respectively.After frozen, recovery, still keep the ability of stably excreting mAb.
(2) the IgG subgroup identification of mAb
Mouse IgG subclass detection kit with HBT company provides is identified hybridoma excretory mAb, and subclass is IgG2b (κ), and tiring reaches 1: 10
4
(3) CHARACTERISTICS IDENTIFICATION of mAb
(3.1) Western blot analyzes
Western blot result shows that hybridoma cell strain hTSC29 excretory mAb of the present invention all can discern M in people's testis tissue specifically
rBe about 60 000 albumen, and with endometrial cell albumen and white blood disease K-562 cellular proteins between do not combine.GAPDH is as confidential reference items, at three kinds of proteic M
rBe that 36 000 places all can detect specific band, as shown in Figure 1.
(3.2) immunohistochemical methods of mAb is identified
Showed by immune group result, the molecule that hTSC29 mAb is discerned mainly is positioned at the spermatocyte and the spermatid of normal people's testis tissue, has explained that hTSC29 albumen has expression in testis, and is as shown in Figure 2.
(3.3) identified by immunofluorescence of mAb
Tangible green fluorescence is arranged in spermatocyte and the spermatid in normal people's testis tissue, painted in the form of a ring, show hTSC29 mAb can with the hTSC29 protein binding in the testis, as shown in Figure 3.
The antibody of the anti-testicle-protein hTSC 29 of the present invention or antibody fragment capable specificity and the protein bound that includes people's testis hTSC29 polypeptide; Antibody or antibody fragment can combine with certain affinity tag such as enzyme, fluorescent marker, chemiluminescent labels, and this antibody or antibody fragment can be applied to Westernblot, ELISA, immunohistochemistry or immunofluorescence dyeing and detect.To those skilled in the art, be easy to learn that this antibody or antibody fragment can also join with the medicine coupling, be used for rake to treatment.
Above content is to combine concrete preferred implementation to the further explain that the present invention did, and can not assert that practical implementation of the present invention is confined to these explanations.For the those of ordinary skill of technical field under the present invention, under the prerequisite that does not break away from the present invention's design, can also make some simple deduction or replace, all should be regarded as belonging to protection scope of the present invention.
Claims (5)
1. hybridoma cell strain hTSC29, its preserving number is CCTCC NO:C200952.
2. one kind by the described hybridoma cell strain hTSC29 of claim 1 excretory antibody.
3. the application of the antibody of an anti-testicle-protein hTSC 29; It is characterized in that: said antibody and coupling matter coupling join; Said coupling matter is enzyme, fluorescent marker or chemiluminescent labels, and said antibody is by the described hybridoma cell strain hTSC29 secretion of claim 1.。
4. the antibody complex of an anti-testicle-protein hTSC 29; It is characterized in that: comprise the antibody of anti-testicle-protein hTSC 29 and the coupling matter that joins with its coupling; Said coupling matter is selected from affinity tag, and said antibody is by the described hybridoma cell strain hTSC29 secretion of claim 1.
5. antibody complex according to claim 4 is characterized in that: said affinity tag is enzyme, fluorescent marker or chemiluminescent labels.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101886409A CN101775371B (en) | 2009-12-02 | 2009-12-02 | Anti-testicle-protein hTSC29 antibody or antibody fragment and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101886409A CN101775371B (en) | 2009-12-02 | 2009-12-02 | Anti-testicle-protein hTSC29 antibody or antibody fragment and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101775371A CN101775371A (en) | 2010-07-14 |
| CN101775371B true CN101775371B (en) | 2012-03-14 |
Family
ID=42511957
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009101886409A Expired - Fee Related CN101775371B (en) | 2009-12-02 | 2009-12-02 | Anti-testicle-protein hTSC29 antibody or antibody fragment and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101775371B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107315092B (en) * | 2017-07-11 | 2019-03-19 | 上海市第一人民医院 | A kind of immunofluorescence staining and its kit of rapid evaluation testicular spermatogenic function |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1352138A (en) * | 2000-11-09 | 2002-06-05 | 中国科学院上海生物工程研究中心 | Sperm formation relative protein and its coding sequence and use |
| CN1459458A (en) * | 2002-05-20 | 2003-12-03 | 卢光琇 | Testes spermatogenous cell die related gene |
| CN1643148A (en) * | 2002-02-14 | 2005-07-20 | 独立行政法人科学技术振兴机构 | Mouse spermatogenesis genes, human male sterility-associated genes and diagnostic system using the same |
| CN1781944A (en) * | 2005-09-01 | 2006-06-07 | 中国人民解放军南京军区南京总医院 | Sperm protein monoclonal antibody and its preparing method and use |
-
2009
- 2009-12-02 CN CN2009101886409A patent/CN101775371B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1352138A (en) * | 2000-11-09 | 2002-06-05 | 中国科学院上海生物工程研究中心 | Sperm formation relative protein and its coding sequence and use |
| CN1643148A (en) * | 2002-02-14 | 2005-07-20 | 独立行政法人科学技术振兴机构 | Mouse spermatogenesis genes, human male sterility-associated genes and diagnostic system using the same |
| CN1459458A (en) * | 2002-05-20 | 2003-12-03 | 卢光琇 | Testes spermatogenous cell die related gene |
| CN1781944A (en) * | 2005-09-01 | 2006-06-07 | 中国人民解放军南京军区南京总医院 | Sperm protein monoclonal antibody and its preparing method and use |
Non-Patent Citations (5)
| Title |
|---|
| Strausberg R.L. |
| Strausberg,R.L.,et al..GenBank: BC032957.1.《NCBI》.2006, * |
| Subbarao S,et al..Genetic characterization of incident HIV type 1 subtype E and B strains from a prospective cohort of injecting drug users in Bangkok, Thailand.《AIDS Res Hum Retroviruses》.2000,第16卷(第8期),699-707. * |
| Subbarao,S.,et al..GenBank: AAC33984.1.《NCBI》.2000, * |
| SubbaraoS et al..Genetic characterization of incident HIV type 1 subtype E and B strains from a prospective cohort of injecting drug users in Bangkok |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101775371A (en) | 2010-07-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105209489B (en) | For the specific detection tool for the circulating tumor cell that interstitial and epithelial-mesenchymal convert | |
| CN101939336B (en) | AXL antibodies | |
| CN107108738A (en) | Anti-cd 3 antibodies, it anti-cd 3 antibodies, polyspecific anti-cd 3 antibodies, polyspecific can be activated can activate anti-cd 3 antibodies and its application method | |
| CN111363043B (en) | anti-CD 20 protein monoclonal antibody, cell line, preparation method and application thereof | |
| CN111410690B (en) | anti-CK 19 protein monoclonal antibody, cell line, preparation method and application thereof | |
| CN104334581B (en) | Anti- c Met antibody | |
| CN105007941B (en) | Anti- LAMP1 antibody and antibody drug conjugates and application thereof | |
| CN101679485B (en) | Osteopontin functional epitopes, monoclonal antibodies against the epitopes and uses thereof | |
| CN106978400A (en) | Anti- PD L1 protein monoclonal antibodies and application thereof | |
| CN105273083A (en) | Anti-human aryl hydrocarbon receptor monoclonal antibody and application thereof | |
| CN111454360A (en) | anti-CD 61 protein monoclonal antibody, cell line, preparation method and application thereof | |
| CN101775371B (en) | Anti-testicle-protein hTSC29 antibody or antibody fragment and application thereof | |
| CN103626871A (en) | Monoclonal antibodies capable of specifically recognizing EGFR mutant protein, preparation method and application thereof | |
| CN102818896B (en) | Detection method of nitration modification sites based on specific antibodies and antibody capable of specifically recognizing succinyl-CoA: 3-oxoacid CoA transferase (SCOT) nitration sites | |
| CN113845592B (en) | anti-CK 5/6 protein monoclonal antibody, cell strain thereof, preparation method and application | |
| CN103172744B (en) | The monoclonal antibody of specific recognition B-Raf mutain, preparation method and application thereof | |
| CN109679924A (en) | The anti-human monoclonal antibody and the preparation method and application thereof with peptide element of high-affinity | |
| CN101921336B (en) | Monoclonal antibody against human alpha1 acid glycoprotein and preparation method thereof | |
| CN111647565B (en) | Preparation method and application of anti-RNF 138 monoclonal antibody | |
| CN103266089A (en) | Anti-human copeptin monoclonal antibody mcco1 and application thereof | |
| CN101775372B (en) | Anti-testicle-protein hT279 antibody or antibody fragment and application thereof | |
| CN105017425B (en) | Anti- HER2 neutralization activities monoclonal antibody and its application | |
| CN106854245A (en) | Protein monoclonal antibody of AntiCD3 McAb 0 and application thereof | |
| CN107043752A (en) | Hybridoma cell strain and the anti-CRH antibody N D19 based on hybridoma cell strain and its application of context of detection | |
| CN103013926B (en) | Anti-cellular-prion-protein monoclonal antibody and application of in diagnostic test kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120314 Termination date: 20131202 |