CN102786595B - Anti-CD5 monoclonal antibody and purpose thereof - Google Patents
Anti-CD5 monoclonal antibody and purpose thereof Download PDFInfo
- Publication number
- CN102786595B CN102786595B CN201210274198.3A CN201210274198A CN102786595B CN 102786595 B CN102786595 B CN 102786595B CN 201210274198 A CN201210274198 A CN 201210274198A CN 102786595 B CN102786595 B CN 102786595B
- Authority
- CN
- China
- Prior art keywords
- monoclonal antibody
- cell
- albumen
- protein
- chip
- 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.)
- Active
Links
Images
Abstract
The invention discloses a hybridoma cell strain (the preservation number is CGMCC No. 6133), and a monoclonal antibody 2B8 derived from the hybridoma cell strain. The invention also relates to the application of the monoclonal antibody 2B8 in preparing an immunity testing tool for detecting the CD5 protein, actually, an immunohistochemical kit containing the monoclonal antibody 2B8, as well as the application of the monoclonal antibody 2B8 in preparing a kit for diagnosing tumors derived from B cells and rheumatoid arthritis. The monoclonal antibody provided by the invention can be combined with the CD5 protein specificity, has no cross reaction with other proteins in the cells, and greatly improves the specificity and reliability of the CD5 protein immunodetection.
Description
Technical field
The present invention relates to can specific combination CD5 albumen monoclonal antibody, and the application in detecting CD5 albumen.
Background technology
CD5 is I type glycoprotein, and is scavenging agent receptor family member.CD5 is expressed by thymocyte, mature T cells and part mature B cell (being mainly the B cell of secretion IgM), and the process of lymphocyte activation and differentiation is relevant with regulating.CD5 antibody contributes to lymphoma to classify, and if jacket cell lymphoma is usually CD5 positive expression, and germinal-center lymphoma is often negative; Derive from the chronic lymphocytic leukemia positive expression of B cell, the lymphoma of other types as follicular lymphoma, hairy cell leukemia, large celllymphoma etc. be that CD5 is negative.Clinically, CD5 can be used as the biomarker of chronic lymphocytic leukemia, lymphoma mantle cell and diffuse large B cell lymphoma.
In addition, there is report to show, rheumatoid arthritis is relevant with the level of the bone-marrow-derived lymphocyte of the CD5 positive: the positive bone-marrow-derived lymphocyte level of rheumatoid arthritis patients CD5 is significantly higher than the positive bone-marrow-derived lymphocyte level of normal healthy controls group CD5, and highly the positive bone-marrow-derived lymphocyte level of movable rheumatoid arthritis patients CD5 is significantly higher than low movable rheumatoid arthritis patients CD5+B lymphocyte level; Therefore, the detection of CD5 expression level can be used for auxiliary diagnosis and the prognosis evaluation of rheumatoid arthritis.
Conventionally by the experiment of immunohistochemistry (IHC) pathology, detect clinically at present the expression situation of CD5 in cell.The core of IHC experiment is the monoclonal antibody of specific binding CD5, and the quality of its performance is directly determining sensitivity and the specificity of whole detection.Therefore, developing a kind of high specific has great importance in conjunction with the anti-CD5 monoclonal antibody of CD5 albumen.
Summary of the invention
The technical problem to be solved in the present invention is for the monoclonal antibody of the anti-CD5 albumen that a kind of binding specificity is higher is provided, and the application in the immunodetection instrument for the preparation of detection CD5 albumen.
For solving the problems of the technologies described above, the invention provides a kind of hybridoma cell strain, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (referred to as CGMCC), preservation date is on May 16th, 2012, and deposit number is CGMCC No.6133.
The present invention also provides a kind of monoclonal antibody 2B8 of specific binding CD5 albumen, by above-mentioned hybridoma cell strain, is produced.
The preparation method of monoclonal antibody of the present invention is as follows:
(1) structure of recombinant expression vector: carry out pcr amplification according to the ORF complete sequence of CD5 gene (as shown in SEQ ID No.1) design primer, restriction endonuclease sites SgfI and MluI are introduced respectively in gene both sides, and introduce Myc-DDK sequence label (as shown in SEQ ID No.2) at its C-terminal, insert expression vector pCMV6-Entry, build CD5 recombinant expression plasmid;
(2) expression and purification of CD5 recombinant protein: with this plasmid transfection HEK293T cell, cracking centrifuging and taking supernatant, is used DDK affinity chromatography column purification, obtains the CD5 recombinant protein of purifying;
(3) screening of monoclonal antibody and preparation: the CD5 recombinant protein immunity BALB/c mouse that adopts above-mentioned purifying, get mouse spleen cell and SP2/0 cell merges, limiting dilution assay obtains mono-clonal, ELISA method screening positive hybridoma cell, obtain the hybridoma cell strain that can secrete anti-CD5 specific antibody, the anti-CD5 antibody called after 2B8 of this hybridoma cell strain secretion, hypotype is accredited as IgG1; Prepare mouse ascites, by affinity chromatography column purification CD5 monoclonal antibody.By Western Blot, immunohistochemical experiment, immunofluorescence, verify respectively sensitivity and the specificity of this monoclonal antibody.
The specificity checking of said monoclonal antibody:
On OriGene high-density protein chip, comprise 10400 HEK293T cell proteins and cross expression lysate, every kind of protein lysate has the repetition of two copies on chip.Protein lysate by trace on nitrocellulose filter.The location of each clock protein lysate can accurately be located by Excel file.On protein chip, albumen is divided into 40 sub-matrixes, on each sub-matrix, has some references, and by reference, quantitatively each chip point is gone up the content of albumen, monitors the repeatability of each immune response data, and the direction of location positive signal.
Monoclonal antibody 2B8 of the present invention and said chip are hybridized and determine positive signal site, result shows: monoclonal antibody 2B8 specific binding CD5 albumen of the present invention, and with other albumen no cross reactions.
The present invention also provides the application of monoclonal antibody 2B8 in the immunodetection instrument for the preparation of detection CD5 albumen.
Particularly, described immunodetection instrument is test kit, chip or test paper.
In specific embodiment, the invention provides a kind of immunologic combined detection reagent kit, comprise said monoclonal antibody 2B 8; Can detect the expression situation of CD5 in histocyte.
The present invention also provides the application of said monoclonal antibody in the test kit for the preparation of diagnosis B cell derived tumour, and described B cell derived tumour is chronic lymphocytic leukemia, lymphoma mantle cell or diffuse large B cell lymphoma.CD5 molecule, as the molecular marked compound of B cell derived tumour, is used monoclonal antibody 2B8 of the present invention to carry out immunodetection to CD5 protein molecular and can be used for diagnosing B cell derived tumour; At present, determined that clinically CD5 molecule is the marker of chronic lymphocytic leukemia, lymphoma mantle cell and diffuse large B cell lymphoma.
In view of the level of the positive B cell of CD5 and the dependency of rheumatoid arthritis, the present invention also provides the application of said monoclonal antibody in the test kit for the preparation of diagnostics classes rheumatic arthritis.
Preservation information
The scientific description that is used for the hybridoma cell strain of preservation is: anti-human CD5 monoclonal antibody hybridoma cell strain 2B8;
Depositary institution's full name: China Committee for Culture Collection of Microorganisms's common micro-organisms center;
Depositary institution is called for short: CGMCC;
Depositary institution address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica;
Preservation date: on May 16th, 2012;
Deposit number: CGMCC No.6133.
Accompanying drawing explanation
Fig. 1 is the expression of Western blot checking CD5 recombinant plasmid in HEK293T cell; The loading sample of left side swimming lane is the HEK293T cell pyrolysis liquid of transfection empty carrier, and the loading sample of the right swimming lane is the HEK293T cell pyrolysis liquid of transfection recombinant plasmid pCMV6-CD5, and hybridizing primary antibodie used is DDK tag antibody;
Fig. 2 is the SDS-PAGE evaluation figure of CD5 recombinant protein;
Fig. 3 is the CD5 expression (take 2B8 monoclonal antibody as primary antibodie) that immunohistochemical methods detects lymphatic cancer tissue;
Fig. 4 is the CD5 expression (take 2B8 monoclonal antibody as primary antibodie) that immunohistochemical methods detects tonsil;
Fig. 5 is the positive signal site of Western blot proofing chip;
Left figure: the 2B8 antibody (1:500 dilution) of take is the results of hybridization of primary antibodie, the 293T cell pyrolysis liquid, albumen Marker, wild-type 293T cell pyrolysis liquid, the 293T cell pyrolysis liquid of excessively expressing CETN2, the mistake that were followed successively by from left to right expression CD5 plasmid are expressed the 293T cell pyrolysis liquid of JUB;
Right figure: take and secrete the results of hybridization that the hybridoma culture supernatant of 2B 8 antibody is primary antibodie, be followed successively by from left to right express CD5 plasmid 293T cell pyrolysis liquid, albumen Marker, wild-type 293T cell pyrolysis liquid, cross express CETN2 293T cell pyrolysis liquid, cross the 293T cell pyrolysis liquid of expressing JUB.
Embodiment
In order to make those skilled in the art understand better technical scheme of the present invention, below in conjunction with specific embodiment, the present invention is described in further detail.
The preparation of the anti-CD5 monoclonal antibody of embodiment 1
One, the structure of CD5 recombinant expression plasmid
The plasmid BC027901(obtaining from ATCC of take contains CD5 ORF sequence shown in SEQ ID No.1) be template, design two primers and introduce respectively restriction enzyme site SgfI, MluI, and C-terminal Myc-DDK label (sequence is as shown in SEQ ID No.2), and be cloned into expression vector pCMV6-Entry, build CD5 recombinant expression plasmid.
Two, the expression and purification of CD5 recombinant protein
1, transfection HEK293T cell
HEK293T cell be take 1:3 and is reached in the Tissue Culture Dish that diameter is 10cm and to continue to cultivate; Get 7.5ml DMEM(serum-free and microbiotic) substratum to 50ml pipe, add 300 μ lPEI MegaTran1.0 to mix, then add the above-mentioned CD5 recombinant plasmid dna of 75 μ g, mix and standing 30 minutes; Get respectively the above-mentioned mixed solution of 515 μ l and add in above-mentioned each Tissue Culture Dish, in 37 ℃, 5%CO
2in incubator, continue to cultivate.After transfection 24 hours, every culture dish cell adds 25 μ l 2M Sodium propanecarboxylates to final concentration 5mM.
2, lysing cell
After transfection 48 hours, carry out lysis.Suck substratum, add 1ml PBS to carry out rinsing, suck PBS.Add 1ml lysis buffer, before use, add proteinase inhibitor PI and PMSF.Be placed in ice chest and vibrate on shaking table, collect the lysate in all culture dish, 4 ℃ centrifugal, collects supernatant.
3, DDK affinity chromatography column purification
With aperture 0.45 μ m, the lysate supernatant that the pvdf membrane filter of diameter 33mm filters after centrifugal also proceeds in 15ml pipe, adds the Sepharose Beads 1ml mixing, and puts into 360 degree vortex mixers after sealing, in 4 ℃ in conjunction with 2 hours; Take out 15ml pipe, lysate is poured in BIO-RAD chromatography column, and catch and penetrate liquid, penetrate liquid sampling WB after dripping to the greatest extent to detect, see Fig. 1; With lysis buffer, rinse post material 1-2 time, after dripping to the greatest extent, with TBST, rinse Beads 3 times again, after dripping to the greatest extent, use 0.1M Glycine(pH3.5) wash-out, 200 μ l for the first time, drip to the greatest extent and do not collect, each 500 μ l of second and third time, the 4th 250 μ l, be collected in a 1.5ml Tube, and add rapidly NaH
2pO
4(pH=11.0) be neutralized to about pH7.0, every pipe adds glycerine to final concentration be 10%, Tween-80 to final concentration be 0.1%.CD5 recombinant protein after purifying is identified with SDS-PAGE, the results are shown in Figure 2.
Three, the screening of monoclonal antibody and preparation
1, animal immune: the CD5 recombinant protein of above-mentioned purifying is with complete Freund's adjuvant emulsification, adopt subcutaneous or abdominal injection method immunity 6-8 BALB/c mouse in age in week, immunizing dose is 50 μ g/, immunity is for the second time carried out at interval after two weeks, with incomplete Freund's adjuvant emulsification, immunizing dose is 50 μ g/.After immune twice, get tail blood and measure serum titer with ELISA method gradient dilution; According to result, determine whether booster immunization, choose the mouse that antibody titer is the highest and carry out cytogamy.
2, cytogamy: myeloma cell adopts the sp2/0 in BALB/c mouse source, during fusion in logarithmic phase; Get the mouse spleen of above-mentioned immunity, make lymphocyte single cell suspension; Immune mouse splenic lymphocyte is mixed with 1:5-1:10 with myeloma cell, drip the 50%PEG(PH 8.0 of 37 ℃) 1ml, add incomplete substratum and stop buffer thereof, centrifugal abandoning adds HAT substratum to suspend after supernatant to mix, MC constant volume is to 50ml, divide and install in 3.5cm culture dish, be put in wet box, be placed in 37 ℃, 5%CO
2in constant incubator, cultivate.
3, screening and clone: merge in 7-10 days and select cell clone, use the CD5 recombinant protein of above-mentioned purifying to carry out ELISA test, labeled cell strain number.Positive porocyte is carried out to limiting dilution, within after each limiting dilution 5-6 days, measure ELISA value, limiting dilution is carried out in the higher mono-clonal hole of the positive value of picking OD280, until ELISA measures 96 orifice plates, entirely hardens really positive; The high mono-clonal of the positive value of picking is determined strain, and its corresponding fusion plate cell strain is 2B8.
4, the male BALB/c mouse abdominal injection 0.5ml pristane in age in the preparation and purification of ascites monoclonal antibody: 10-12 week, one week afterwards every mouse with 1ml syringe abdominal injection, through PBS, wash resuspended monoclonal cell suspension, cell consumption is 5 * 10
6/ only, every strain antibody is made a call to 2 mouse.After gathering, collects mouse ascites ascites, centrifuging and taking supernatant, and affinity chromatography carries out ascites purifying, according to antibody subtype, selects respective post material, and monoclonal antibody 2B 8 hypotypes are IgG1, adopt proteinG to carry out purifying.Monoclonal antibody concentration determination after purifying, packing, frozen at-20 ℃.
Embodiment 2 be take monoclonal antibody 2B8 and lymphatic cancer tissue is carried out to immunohistochemical methods detection as primary antibodie
1, get lymphatic cancer tissue and carry out paraffin embedding, use Finesse histotome to cut into slices, tissue thickness is 6 μ m;
2, dewaxing and aquation: 3 times * 10min of analytical pure dimethylbenzene, 3 times * 10min of dehydrated alcohol, 95% ethanol 5min, 85% ethanol 5min, 75% ethanol 5min, deionized water soaks 3min * 3 time;
3, add antigen retrieval liquid (0.01M, pH6.0 sodium citrate buffer) pressure kettle hot high pressure to repair 2min, when pressure kettle temperature is down to approximately 90 ℃, open pressure kettle, take out sample, then naturally cool to room temperature.Deionized water soaks 3min * 3 time.
4, use 3% hydrogen peroxide deactivation to organize endogenous peroxydase, the standing 10min of room temperature.Deionized water soaks 5min * 3 time.
5, add confining liquid (PBS+5% skim-milk+5% normal goats serum), hatch 60min for 37 ℃.
6, remove confining liquid, do not rinse, add the monoclonal antibody 2B8 of the present invention of 1:150 dilution proportion; Be placed in wet box, hatch 60min for 37 ℃.PBST(is containing 0.1%Tween-20) wash 2 times, wash 5min at every turn.PBST(is containing 0.02%Tween-20) wash 1 time, wash 5min at every turn.
7, drip Polink-test kit 2(Catlog No.D37-15) in 1,37 ℃ of reagent hatch 10-20 minute.Use PBS washing 3 times, each 5min.Drip 2,37 ℃ of reagent in Polink-2 test kit (Catlog No.D37-15) and hatch 10-20 minute, use PBS washing 3 times, each 5min.
8, application DAB solution (ZLI-9019 of Zhong Shan Golden Bridge) colour developing, colour developing 3 ~ 10min.Distilled water wash.
9, haematoxylin redyeing nucleus 2min, distilled water rinsing, 1% hydrochloric acid differentiation.Distilled water rinsing 3 times, the standing 1min of room temperature.
10, dehydration and transparent: 75% ethanol 5min, 100% ethanol 5min * 3 time, 85% ethanol 5min, 95% ethanol 5min, 100% ethanol 3 * 5min; Dimethylbenzene 3 * 5min, neutral gum mounting.
11, microscopy, is shown in Fig. 3.
As seen from Figure 3: the visible a large amount of brown yellow granules of lymphatic cancer tissue are CD5 protein expression positive cell.
Embodiment 3 be take monoclonal antibody 2B8 and tonsil is carried out to immunohistochemical methods detection as primary antibodie
Use 8 pairs of tonsils of monoclonal antibody 2B of the present invention to carry out immunohistochemical methods detection, detecting step is with embodiment 2, and as shown in Figure 4, as seen from Figure 4, the visible a large amount of brown yellow granules of tonsil, are CD5 protein expression positive cell to result.
Embodiment 4, OriGene protein chip detect the specificity of monoclonal antibody 2B8
On OriGene high-density protein chip, comprise 10400 HEK293T cell proteins and cross expression lysate, every kind of protein lysate has the repetition of two copies on chip.Protein lysate by trace on nitrocellulose filter.The location of each clock protein lysate can accurately be located by Excel file.On protein chip, albumen is divided into 40 sub-matrixes, on each sub-matrix, has some references, and by reference, quantitatively each chip point is gone up the content of albumen, monitors the repeatability of each immune response data, and the direction of location positive signal.
Below for using OriGene albumen (OriGene Cat PA100001) chip 2B8 monoclonal antibody to be carried out to the concrete steps of specificity identification:
1, a protein chip is placed in 50ml centrifuge tube, uses 40ml deionized water to infiltrate chip, be placed on shaking table mixed at room temperature 30 minutes; Discard deionized water, use 10mlPBST balance chip, room temperature treatment 10 minutes.
2, in 50ml centrifuge tube, add 40ml 5% skimmed milk (diluting with PBST) to be placed on shaking table, room temperature sealing 30 minutes.
3, use confining liquid (5% skimmed milk) dilution primary antibodie 2B 8, Dilution ratio 1:200.
4, clean sealed membrane is pasted on experiment table, drip the above-mentioned dilution primary antibodie of 250-300 μ l on sealed membrane.
5, protein chip is extracted out from confining liquid, faced down one of protein chip NC film, one side contact antibody from chip, slowly slides, and relies on surface tension of liquid, and antibody will slowly infiltrate chip NC film, until whole NC film infiltrates in primary antibodie solution.Whole operating process avoids producing bubble.Chip is moved on under 4 degree environment, standing, primary antibodie overnight incubation.On chip, add a cover culture dish lid, on it, stick a hygenic towelette, to prevent from hatching for a long time, cause antibody to evaporate.
6, second day moves to chip in 50ml centrifuge tube, uses PBST rinsing chip twice, removes unnecessary antibody.Use 40ml PBST(0.1%Tween-20) washing chip, be placed on shaking table and mix, wash three times, wash 5min at every turn.
7, use confining liquid (5% skimmed milk) dilution two anti-DyLight649-conjugated AffiniPure Fragment Goat-anti-Mouse IgG, Dilution ratio is 1:400.
8,, according to above-mentioned steps 4, step 5 is carried out two and anti-is hatched operation.Incubated at room 1 hour.Above chip, with aluminium-foil paper, hide, to prevent signal bleaching.
9,, according to above-mentioned steps 6, use PBST washing chip.
10, use deionized water rinsing chip, to remove remaining salinity and denaturing agent.
11, drying at room temperature chip, guarantees chip complete drying.
12, use chip scanner to read fluorescent signal.
13, according to BSA-Cy3 and BSA-Cy5, determine the site of chip direction and positive signal.
14, according to positive signal site, find out corresponding protein lysate ID, according to lysate database information, find corresponding protein name, NCBI typing number (accession number), protein I D, the information such as albumen size.
Chip results shows: on protein chip, occur 4 positive signal points, corresponding albumen is respectively CD5, CETN2, JUB, CACNG5.
Embodiment 5, Western blot checking protein chip results of hybridization
Use Western blot to carry out analysis verification to embodiment 4 acquired results.To cross cell pyrolysis liquid and the wild-type 293T cell pyrolysis liquid of the 293T cell of expressing CD5 recombinant plasmid, CETN2 recombinant plasmid, JUB recombinant plasmid, carry out loading respectively, after SDS-PAGE electrophoresis, transferring film, sealing, using respectively the hybridoma supernatant of 2B 8 monoclonal antibodies of the present invention (Dilution ratio is 1:500) and secretion 2B8 antibody is that primary antibodie is hybridized, and its result as shown in Figure 5.
As seen from Figure 5: no matter be 2B8 monoclonal antibody of the present invention or the Hybridoma Cell Culture supernatant of secretion 2B8 antibody, all only with CD5 protein binding, and with CETN2, JUB proteantigen all without combination, prompting: monoclonal antibody 2B8 of the present invention is specific binding CD5 albumen only.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (8)
1. a monoclonal antibody 2B8 for specific binding CD5 albumen, the hybridoma cell strain that is CGMCC No.6133 by deposit number produces.
2. a hybridoma cell strain, its deposit number is CGMCC No.6133.
3. the application of monoclonal antibody as claimed in claim 1 in the immunodetection instrument for the preparation of detection CD5 albumen.
4. application according to claim 3, is characterized in that, described immunodetection instrument is test kit, chip or test paper.
5. an immunologic combined detection reagent kit, is characterized in that, comprises monoclonal antibody claimed in claim 1.
6. monoclonal antibody as claimed in claim 1 is being usingd CD5 albumen as the application in the B cell derived tumor reagent box of molecular marked compound for the preparation of diagnosis.
7. application according to claim 6, is characterized in that, described to take CD5 albumen be chronic lymphocytic leukemia, lymphoma mantle cell or diffuse large B cell lymphoma as the B cell derived tumour of molecular marked compound.
8. the application of monoclonal antibody as claimed in claim 1 in the rheumatoid arthritis test kit that positive B cell high level is relevant for the preparation of diagnosis CD5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210274198.3A CN102786595B (en) | 2012-08-03 | 2012-08-03 | Anti-CD5 monoclonal antibody and purpose thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210274198.3A CN102786595B (en) | 2012-08-03 | 2012-08-03 | Anti-CD5 monoclonal antibody and purpose thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102786595A CN102786595A (en) | 2012-11-21 |
| CN102786595B true CN102786595B (en) | 2014-04-23 |
Family
ID=47152199
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210274198.3A Active CN102786595B (en) | 2012-08-03 | 2012-08-03 | Anti-CD5 monoclonal antibody and purpose thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102786595B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20240050568A1 (en) * | 2021-01-12 | 2024-02-15 | Nanjing IASO Biotechnology Co., Ltd. | Fully human single-domain tandem chimeric antigen receptor (car) targeting cd5, and use thereof |
| CN113105547B (en) * | 2021-05-18 | 2022-04-08 | 福州迈新生物技术开发有限公司 | anti-CD 5 protein monoclonal antibody and cell strain, preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1351259A (en) * | 2000-10-31 | 2002-05-29 | 杨梦甦 | Protein chip for immunodiagnosis and its preparing process |
| CN1508155A (en) * | 2002-12-18 | 2004-06-30 | 菁 马 | Anti CD52 monoclonal antibody, coding sequence and use thereof |
| CN102137873A (en) * | 2008-08-29 | 2011-07-27 | 西福根有限公司 | Anti-CD5 antibodies |
| CN102245783A (en) * | 2008-10-08 | 2011-11-16 | 剑桥企业有限公司 | Methods and compositions for diagnosis and treatment |
-
2012
- 2012-08-03 CN CN201210274198.3A patent/CN102786595B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1351259A (en) * | 2000-10-31 | 2002-05-29 | 杨梦甦 | Protein chip for immunodiagnosis and its preparing process |
| CN1508155A (en) * | 2002-12-18 | 2004-06-30 | 菁 马 | Anti CD52 monoclonal antibody, coding sequence and use thereof |
| CN102137873A (en) * | 2008-08-29 | 2011-07-27 | 西福根有限公司 | Anti-CD5 antibodies |
| CN102245783A (en) * | 2008-10-08 | 2011-11-16 | 剑桥企业有限公司 | Methods and compositions for diagnosis and treatment |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102786595A (en) | 2012-11-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102827281B (en) | Monoclonal antibody against CD2 protein and application thereof | |
| CN106978400A (en) | Anti- PD L1 protein monoclonal antibodies and application thereof | |
| CN106188292B (en) | Anti- CD4 protein monoclonal antibody and application thereof | |
| CN105254759A (en) | Anti-CD56 protein monoclonal antibody hybridoma cell, anti-CD56 monoclonal antibody generated by same and application | |
| CN102924594B (en) | Anti-tumor-protein-P63 monoclonal antibody and purpose thereof | |
| CN105131114B (en) | The anti-p16 monoclonal antibodies and application of anti-p16 protein monoclonal antibodies hybridoma and its generation | |
| CN102786595B (en) | Anti-CD5 monoclonal antibody and purpose thereof | |
| CN103102414B (en) | HER2-resistant protein monoclonal antibody and application thereof | |
| CN111848750B (en) | Method and kit for rapidly enriching and detecting 2019-nCoV | |
| CN105175543A (en) | E-Cadherin protein resisting monoclonal antibody hybridoma, E-Cadherin resisting monoclonal antibody generated by same and application of E-Cadherin resisting monoclonal antibody | |
| CN110357964A (en) | 1 protein monoclonal antibody of AntiCD3 McAb and application thereof | |
| CN107446046A (en) | Anti- CD20 protein monoclonal antibodies and application thereof | |
| CN105622751B (en) | Anti- Ki67 protein monoclonal antibody and application thereof | |
| CN106749668A (en) | Anti- IDO1 protein monoclonal antibodies and application thereof | |
| CN102827278B (en) | Antitumor protein P53 monoclonal antibody and application thereof | |
| CN109096400A (en) | Anti- PDPN protein monoclonal antibody and application thereof | |
| CN106977605A (en) | Anti- CD19 protein monoclonal antibodies and application thereof | |
| CN105238763A (en) | Anti-PR protein monoclonal antibody hybridomas cell, anti- PR monoclonal antibody generated by same and application | |
| CN106854245A (en) | Protein monoclonal antibody of AntiCD3 McAb 0 and application thereof | |
| CN107236040A (en) | Anti- MSH6 protein monoclonal antibodies and application thereof | |
| CN105237638A (en) | Anti-EGFR (epidermal growth factor receptor) protein monoclonal antibody hybridomas cell, anti-EGFR monoclonal antibody generated by same and application | |
| CN102911271B (en) | Anti-CTNNB1 monoclonal antibody 12H7 and application thereof | |
| CN102898520B (en) | Anti-CTNNB1 monoclonal antibody 3F9 and application thereof | |
| CN107043419A (en) | Anti- BTLA protein monoclonal antibodies and application thereof | |
| CN109111519A (en) | Anti- S100P protein monoclonal antibody and application thereof |
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 |