CN105254759A - Anti-CD56 protein monoclonal antibody hybridoma cell, anti-CD56 monoclonal antibody generated by same and application - Google Patents
Anti-CD56 protein monoclonal antibody hybridoma cell, anti-CD56 monoclonal antibody generated by same and application Download PDFInfo
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Abstract
The invention relates to the field of biotechnology, discloses a hybridoma cell strain (the preservation serial number is CGMCC No.11081) and a monoclonal antibody Umab83 generated by the hybridoma cell strain, and further relates to application of the monoclonal antibody Umab83 in preparation of a an immunological detection tool used for detecting CD56 protein, an immunological histochemical reagent kit containing the monoclonal antibody Umab83 and application of the monoclonal antibody Umab83 in preparation of a reagent kit used for making tumors. The monoclonal antibody can be combined with specificity of the CD56 protein and has no cross reaction with other proteins in the cell, and therefore the specificity, accuracy and reliability of CD56 protein immunological detection are remarkably improved.
Description
Technical field
The present invention relates to biological technical field, the anti-CD56 monoclonal antibody of particularly a kind of anti-CD56 protein monoclonal antibody hybridoma and generation thereof and application.
Background technology
CD56 is also called nerve cell adhesion molecule (N-CAM), belongs to I type transmembrane glycoprotein.Extensively be present in the cell of most neuroectodermal origin, tissue and tumour, comprise retinoblastoma, medulloblastoma, astrocytoma, neuroblastoma, small cell carcinoma, be also expressed in tumour, natural killer cell and NK lymphoma that some mesoderms are derivative.
The expression amount of CD56 can reflect the transfer ability of tumour cell, is usually detected the expression situation of CD56 albumen in tumour cell at present clinically by immunohistochemistry (IHC) Pathological experiment.The core of IHC experiment is the monoclonal antibody of specific binding CD56 albumen, and the quality of its performance directly decides sensitivity and the specificity of whole detection.Therefore, the monoclonal antibody for CD56 albumen developing a kind of binding specificity higher has great importance.
Summary of the invention
In view of this, the invention provides anti-CD56 monoclonal antibody and the application of a kind of anti-CD56 protein monoclonal antibody hybridoma and generation thereof.Monoclonal antibody of the present invention can be combined with CD56 protein-specific, and with other albumen no cross reactions in cell, significantly improve specificity, accuracy and reliability that CD56 protein immunization detects, CD56 protein expression level in true reflection tumour cell, can be applicable to auxiliary diagnosis and the somatotype of retinoblastoma, medulloblastoma, astrocytoma, neuroblastoma, small cell carcinoma, NK lymphoma and natural killer cell.
In order to realize foregoing invention object, the invention provides following technical scheme:
The invention provides a kind of monoclonal antibody, it is combined with CD56 protein-specific.
In specific embodiments more of the present invention, the hybridoma cell strain that monoclonal antibody is CGMCCNo.11081 by deposit number produces.
Present invention also offers the application of described monoclonal antibody in the immunodetection instrument for the preparation of detection CD56 albumen.
In specific embodiments more of the present invention, described immunodetection instrument is reagent, test kit, chip or test paper.
Present invention also offers a kind of immunologic combined detection reagent kit, comprise described monoclonal antibody.
Present invention also offers described monoclonal antibody for the preparation of the application in the reagent of tagged tissue cell or test kit.
In specific embodiments more of the present invention, histocyte described in described immunologic combined detection reagent kit is natural killer (NK) cell, NK sample T cell, nerve, neuroendocrine tissue or related neoplasms.Described related neoplasms is and above-mentioned cell, tumour that organization type is identical.
Present invention also offers a kind of test kit of tagged tissue cell, comprise described monoclonal antibody.
In specific embodiments more of the present invention, histocyte described in the test kit of described tagged tissue cell is natural killer (NK) cell, NK sample T cell, nerve, neuroendocrine tissue or related neoplasms.Described related neoplasms is and above-mentioned cell, tumour that organization type is identical.
Present invention also offers a kind of hybridoma cell strain, its deposit number is CGMCCNo.11081.
Compared with prior art, the invention provides a kind of hybridoma cell strain (deposit number is CGMCCNo.11081), and the monoclonal antibody Umab83 of hybridoma cell strain generation thus.Present invention also offers the application of monoclonal antibody Umab83 in the immunodetection instrument for the preparation of detection CD56 albumen, containing the immunohistochemical kit of monoclonal antibody Umab83, and monoclonal antibody Umab83 is for the preparation of the application in the test kit of marked tumor.Monoclonal antibody of the present invention can be combined with CD56 protein-specific, and with other albumen no cross reactions in cell, significantly improve specificity, accuracy and reliability that CD56 protein immunization detects, CD56 protein expression level in true reflection tumour cell, can be applicable to auxiliary diagnosis and the somatotype of retinoblastoma, medulloblastoma, astrocytoma, neuroblastoma, small cell carcinoma, NK lymphoma and natural killer cell.
Biological deposits explanation
Classification And Nomenclature for the hybridoma cell strain UMAB83 of preservation is: the strain of nerve cell adhesion molecule 1 (CD56) monoclonal antibody hybridoma cell;
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 07 03rd, 2015;
Deposit number: CGMCCNo.11081.
Accompanying drawing explanation
In order to be illustrated more clearly in the embodiment of the present invention or technical scheme of the prior art, be briefly described to the accompanying drawing used required in embodiment or description of the prior art below.
Fig. 1 shows that the design of embodiment 1 cloning site is as figure, and wherein shading part is ORF district;
Fig. 2 shows embodiment 2 recombinant C D56 albumen Westernblot detected result figure, detect the expression of recombinant C D56 albumen in HEK293T cell with anti-DDK, wherein the detected result of left side swimming lane to be the HEK293T cell pyrolysis liquid of transfection empty carrier be antigen, right lanes are the detected result of the HEK293T cell pyrolysis liquid antigen of transfection pCMV6-rCD56 plasmid;
Fig. 3 shows embodiment 2 recombinant C D56 protein SDS-PAGE result figure, and with anti-DDK affinity column purification of Recombinant CD56 albumen, the albumen after purifying is by SDS-PAGE glue electricity arteries and veins, coomassie brilliant blue staining;
Fig. 4 shows that embodiment 3 identifies the Westernblot detected result figure of complete CD56 (FulllengthCD56, CD56-FL) albumen with monoclonal antibody Umab83.Swimming lane 1 is the cell pyrolysis liquid of transfection pCMV6-Entry; Swimming lane 2 is the cell pyrolysis liquid of transfection pCMV6-CD56-FL;
Fig. 5 shows that embodiment 4 formalin is fixed, paraffin-embedded grownup's cerebral tissue ImmunohistochemistryResults Results figure (primary antibodie is CD56 monoclonal antibody Umab83);
Fig. 6 shows that embodiment 4 formalin is fixed, paraffin-embedded Human embryo cerebellar tissue ImmunohistochemistryResults Results figure (primary antibodie is CD56 monoclonal antibody Umab83);
Fig. 7 shows that (primary antibodie is CD56 monoclonal antibody Umab83,1:100 to embodiment 5OriGene protein chip qualification result figure; Two resist for DyLight649-conjugatedAffiniPureFragmentGoat-anti-MouseIg G, 1:400).
Embodiment
The invention discloses anti-CD56 monoclonal antibody and the application of a kind of anti-CD56 protein monoclonal antibody hybridoma and generation thereof, those skilled in the art can use for reference present disclosure, and suitable improving technique parameter realizes.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.Method of the present invention and application are described by preferred embodiment, related personnel obviously can not depart from content of the present invention, spirit and scope methods and applications as herein described are changed or suitably change with combination, realize and apply the technology of the present invention.
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 07 03rd, 2015, and deposit number is CGMCCNo.11081.
Present invention also offers a kind of monoclonal antibody Umab83 of specific binding CD56 albumen, produced by above-mentioned hybridoma cell strain.
The preparation method of monoclonal antibody of the present invention is as follows:
(1) structure of recombinant expression vector: according to CD56ORF nucleotide sequence (CD56ORF nucleotide sequence as shown in SEQIDNO.1,2574bp; CD56 aminoacid sequence is as shown in SEQIDNO.2)
Design primer PCR amplification CD56ORF 58bp position is to 2154bp bit sequence, gene both sides introduce restriction endonuclease sites SgfI and MluI respectively, insert expression vector pCMV6-Entry, build the recombinant expression plasmid pCMV6-rCD56 of CD56 aminoacid sequence the 20th to the 718th; Upstream amplification primer sequence, SEQIDNO.3:CAC
gCGATCGCaTGCTGCAGGTGGATATTGTTC downstream amplification primer sequence SEQIDNO.4:ACCG
aCGCGTcACCAGGAGCAGGACGAAGAT
(2) expression and purification of CD56 recombinant protein: CD56 recombinant expression plasmid is transformed HEK293T cell, the centrifugal acquisition supernatant of cracking, DDK affinity chromatography column purification, obtains the CD56 recombinant protein of purifying;
(3) screening of monoclonal antibody and preparation: the CD56 recombinant protein immunity Balb/c mouse adopting above-mentioned purifying, get Mouse spleen cells and SP2/0 cell merges, limiting dilution assay obtains mono-clonal, ELISA method screening positive hybridoma cell, obtain the hybridoma cell strain secreting anti-CD56 specific antibody, called after Umab83, hypotype is accredited as IgG1; By serum free medium Dispersal risk, obtain CD56 monoclonal antibody Umab83 by affinity chromatography column purification.Sensitivity and the specificity of this monoclonal antibody is verified respectively by WesternBlot, immunohistochemical experiment.
The specificity of further employing OriGene high-density protein chip to said monoclonal antibody is verified: OriGene high-density protein chip comprises 10,000 HEK293T cell protein process LAN lysate, often 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 accurately can be located by Excel file.On protein chip, albumen is divided into 40 sub-matrixes, and each sub-matrix has some references, by referring to, can the content of albumen on quantitative each chip point, monitor the repeatability of each immune response data, and the direction of location positive signal.
Described monoclonal antibody Umab83 and said chip are hybridized and are determined positive signal site by the present invention, and result shows monoclonal antibody Umab83 specific binding CD56 albumen of the present invention, and with other albumen no cross reactions.
Present invention also offers the application of monoclonal antibody Umab83 in the immunodetection instrument for the preparation of detection CD56 albumen.
Particularly, described immunodetection instrument is test kit, chip or test paper.
In the particular embodiment, the invention provides a kind of immunologic combined detection reagent kit, comprise said monoclonal antibody Umab83, the expression situation of CD56 in histocyte can be detected.
Present invention also offers said monoclonal antibody for the preparation of the application in the test kit of marked tumor.Wherein said tumour specifically refers to the propagation of tumour cell and the closely-related tumour of expression of CD56, includes but not limited to retinoblastoma, medulloblastoma, astrocytoma, neuroblastoma, small cell carcinoma, NK lymphoma and natural killer cell.
Compared with prior art, the invention provides a kind of hybridoma cell strain (deposit number is CGMCCNo.11081), and the monoclonal antibody Umab83 of hybridoma cell strain generation thus.Present invention also offers the application of monoclonal antibody Umab83 in the immunodetection instrument for the preparation of detection CD56 albumen, containing the immunohistochemical kit of monoclonal antibody Umab83, and monoclonal antibody Umab83 is for the preparation of the application in the test kit of marked tumor.Monoclonal antibody of the present invention can be combined with CD56 protein-specific, and with other albumen no cross reactions in cell, significantly improve specificity, accuracy and reliability that CD56 protein immunization detects, CD56 protein expression level in true reflection tumour cell, can be applicable to auxiliary diagnosis and the somatotype of retinoblastoma, medulloblastoma, astrocytoma, neuroblastoma, small cell carcinoma, NK lymphoma and natural killer cell.
The invention provides monoclonal antibody and uses thereof, produce the hybridoma cell strain of described monoclonal antibody, all can be buied by market containing raw materials used in the diagnostic tool of this monoclonal antibody and reagent.
Below in conjunction with embodiment, set forth the present invention further:
The structure of embodiment 1, CD56 recombinant expression plasmid
With the plasmid RC207890 (containing CD56ORF2574bp) obtained from bio tech ltd of Aureal Dongyuan County of the U.S. for template, design two primers and introduce restriction enzyme site SgfI and MluI respectively, be cloned into expression vector pCMV6-Entry, set up CD56 recombinant expression plasmid.Cloning site design as shown in Figure 1.
The expression and purification of embodiment 2, CD56 recombinant protein
1, transfection HEK293T cell: HEK293T cell reaches in culture dish with 1:3 and continues to cultivate; Get 7.5mLDMEM (serum-free and microbiotic) in 50mL pipe, add 300 μ LPEIMegaTran1.0 and mix; Add 75 μ gCD56 recombinant expression plasmid DNA in mixing liquid, mix and leave standstill 30 minutes; Get respectively in 515 μ L to each culture dish in 37 DEG C of 5%CO
2cultivate in incubator.Transfection is after 24 hours, and every ware cell adds 25 μ L2M Sodium propanecarboxylates to final concentration 5mM.
2, lysing cell: transfection, after 48 hours, carries out lysis.Suck substratum, add 1mLPBS and carry out rinsing, suck PBS.Add 1mL lysis buffer, before using, add proteinase inhibitor PI and PMSF.Be placed in ice chest to vibrate on shaking table, collect in all culture dish and obtain lysate, 4 DEG C centrifugal, collects supernatant.The supernatant that takes a morsel adopts WB to identify the expression of recombinant C D56, result Fig. 2.
3, DDK affinity chromatography column purification: with 0.45 μM, the lysate supernatant after 33mmPVDF membrane filter is centrifugal also proceeds to 15mL pipe, adds the Beads1mL mixed, puts into 360 degree of vortex mixers after sealing, in 4 DEG C in conjunction with 2 hours; Take out 15mL pipe, lysate is poured in BIO-RAD chromatography column, and catch and penetrate liquid, drip and penetrate liquid sampling WB to the greatest extent and detect; Rinse post material 1-2 time with lysis buffer, drip to the greatest extent and rinse Beads3 time with TBST again, drip to the greatest extent and use 0.1MGlycinepH3.5 wash-out, first time 200 μ L, drip and do not collect to the greatest extent, second and third each 500 μ L, third time 250 μ L, are collected in a 1.5mL centrifuge tube, and add NaH rapidly
2pO
4(pH=11.0) be neutralized to about pH7.0, often pipe adds glycerine to final concentration be 10%, Tween-80 is 0.1% to final concentration.Recombinant C D56 albumen SDS-PAGE after purifying identifies, sees Fig. 3.
From Fig. 2 result, in the HEK293T cell pyrolysis liquid of transfection pCMV6-rCD56 plasmid, WB has obvious specific band at 100kD place after detecting, and substantially conforms to the theoretical molecular 80kD of polypeptide.Show recombinant C D56 albumen specifically expressing in cell.
From Fig. 3 result, the albumen of purifying has obvious specific band at PAGE glue 100kD place, substantially conforms to the theoretical molecular 80kD of polypeptide.Show to obtain the good CD56 recombinant protein of purity.
The preparation of embodiment 3, CD56 monoclonal antibody and screening
CD56 protein fragments according to the purifying of standard method restructuring generation is used for carrying out immunity to Balb/c mouse (Beijing Vital River Experimental Animals Technology Co., Ltd.).Concrete grammar is as follows:
1, animal immune: purified CD56 antigen 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/, and second time immunity is carried out at interval after two weeks, with incomplete Freund's adjuvant emulsification, immunizing dose is 50 μ g/.Get tail blood after immune twice and measure serum titer with ELISA method gradient dilution; Determine whether booster immunization according to result, choose the highest mouse of antibody titer and carry out cytogamy.
2, cytogamy: the sp2/0 that myeloma cell adopts Balb/c to originate, is in logarithmic phase during fusion; Get immune mouse spleen, make lymphocyte single cell suspension; Mouse spleen lymphocyte mixes with 1:5-1:10 with myeloma cell, drip 50%PEG (PH8.0) 1mL of 37 DEG C, add incomplete substratum and all the other stop buffers, centrifugal abandon supernatant after add HAT substratum suspend mixing, MC constant volume is to 50mL, be dispensed in 3.5cm culture dish, be put in wet box, be placed in 37 DEG C, 5%CO
2cultivate in constant incubator.
3, screening and clone: merge in 7-10 days and select cell clone, uses CD56 purification of recombinant proteins to carry out ELISA test.Labeled cell strain number.Carry out limiting dilution to positive porocyte, within after each limiting dilution 5-6 days, measure ELISA value, limiting dilution is carried out in the mono-clonal hole that the positive value of picking OD280 is higher, until ELISA measures 96 orifice plates entirely harden fruit for positive.The high mono-clonal of picking positive value determines strain.Its corresponding fusion plate cell strain is Umab83.
4, the male Balb/c mouse peritoneal injection 0.5ml pristane in the preparation and purification of ascites monoclonal antibody: 10-12 age in week, one week afterwards every mouse 1mL syringe abdominal injection wash resuspended monoclonal cell suspension through PBS, cell consumption is 5 × 10
6/ only, every strain antibody makes a call to 2 mouse.After mouse ascites gathers, collect ascites, centrifuging and taking supernatant, affinity chromatography carries out ascites purifying, selects corresponding post material according to antibody subtype, and the monoclonal antibody that cell strain Umab83 produces is IgG1, adopts proteinG to carry out purifying.Monoclonal antibody concentration determination after purifying, WB detection, packing, frozen at-20 DEG C.Wherein WB detected result is shown in Fig. 4.
From Fig. 4 result, swimming lane 2 is about 100kD place at molecular weight special band, shows that monoclonal antibody Umab83 can detect complete total length CD56 albumen by Westernblot specifically.
Embodiment 4, monoclonal antibody Umab83 are that the immunohistochemical methods of primary antibodie detects
(1), experimental technique:
1, the adult brain's tissue getting formalin fixing carries out paraffin embedding with Human embryo cerebellar tissue block, and use Finesse histotome to cut into slices, tissue thickness is 6 μm.
2, dewaxing and aquation: analytical pure dimethylbenzene 3 × 10min, dehydrated alcohol 3 × 10min, 95% ethanol 5min, 85% ethanol 5min, 75% ethanol 5min, deionized water soaks 3min × 3 time
3, add antigen retrieval buffers (0.01M, pH6.0 sodium citrate buffer) pressure kettle hot high pressure and repair 3min, when pressure kettle temperature is down to about 90 DEG C, 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, room temperature leaves standstill 10min.Deionized water soaks 5min × 3 time.
5, add confining liquid (PBS+5% skim-milk+5% Normal Goat Serum), hatch 60min for 37 DEG C.
6, remove confining liquid, do not rinse, add CD56 monoclonal antibody (Umab83), thinning ratio: 1:150, use confining liquid to dilute.Be placed in wet box, hatch 60min for 37 DEG C.PBST (0.1%Tween-20) washs 2 times, washs 5min at every turn.PBST (0.02%Tween-20) washs 1 time, washs 5min at every turn.
7, drip Polink-test kit 2 (CatlogNo.D37-15) reagent 1,37 DEG C and hatch 10-20 minute.PBS is used to wash 3 times, each 5min.Drip Polink-2 test kit (CatlogNo.D37-15) reagent 2,37 DEG C and hatch 10-20 minute, use PBS to wash 3 times, each 5min.
8, DAB solution (Zhong Shan Golden Bridge ZLI-9019) colour developing is applied, colour developing 3 ~ 10min.Distilled water wash.
9, haematoxylin redyeing nucleus 2min, distilled water rinsing, 1% hydrochloric acid differentiation.Distilled water rinsing 3 times, room temperature leaves standstill 1min.
10, dehydration and transparent: 75% ethanol 5min, 100% ethanol 5minx3 time, 85% ethanol 5min, 95% ethanol 5min, 100% ethanol 3 × 5min; Dimethylbenzene 3 × 5min, neutral gum mounting.
11, microscopy, is shown in Fig. 5-6.
(2), experimental result:
From Fig. 5-6 result, specific membranes can be seen and dye in adult brain's tissue with Embryonic Cerebellum tissue.Result and CD56 intracellular location and tissue expression specificity consistent, show that monoclonal antibody Umab83 can be used for the level of Immunohistochemical detection CD56 albumen.
The specific detection of embodiment 5, monoclonal antibody Umab83
OriGene high-density protein chip comprises 10,000 HEK293T cell protein process LAN lysate, often 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 accurately can be located by Excel file.On protein chip, albumen is divided into 40 sub-matrixes, and each sub-matrix has some references, by referring to, can the content of albumen on quantitative each chip point, monitor the repeatability of each immune response data, and the direction of location positive signal.Experimental technique below for using OriGene albumen (OriGeneCatPA100001) chip to carry out the experiment of Umab83 Identification of the antibodies:
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 to balance chip.Room temperature treatment 10 minutes.
2, in 50mL centrifuge tube, adding 40mL5% skimmed milk (diluting with PBST) is placed on shaking table, and room temperature closes 30 minutes.
3, use confining liquid (5% skimmed milk) to dilute primary antibodie Umab83, thinning ratio is classified as 1:100.
4, the sealed membrane of cleaning is pasted on experiment table, drip 250-300 μ L primary antibodie on sealed membrane.
5, extracted out from confining liquid by protein chip, face down one of protein chip NC film, from the contact antibody of chip, slowly slide, rely on surface tension of liquid, antibody slowly will infiltrate chip NC film, until whole NC film infiltrates in primary antibodie solution.Whole operating process is avoided producing bubble.Under chip being moved on to 4 DEG C of environment, leave standstill, primary antibodie overnight incubation.Chip is added a cover culture dish lid, it sticks a hygenic towelette, cause antibody to evaporate to prevent from hatching for a long time.
6, chip was moved in 50mL centrifuge tube in second day, use PBST rinsing chip twice, remove unnecessary antibody.Use 40mLPBST (0.1%Tween-20) to wash chip, be placed on shaking table and mix, wash three times, wash 5min at every turn.
7, use confining liquid (5% skimmed milk) to dilute two anti-DyLight649-conjugatedAffiniPureFragmentGoat-anti-MouseIg G, 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.Hide with aluminium-foil paper above chip, to prevent signal bleaching.
9, according to above-mentioned steps 6, PBST is used to wash chip.
10, deionized water rinsing chip is used, to remove remaining salinity and denaturing agent.
11, drying at room temperature chip, guarantees chip complete drying.
12, chip scanner is used to read fluorescent signal.
13, the site of chip direction and positive signal is determined according to BSA-Cy3 and BSA-Cy5.
14, find out corresponding protein lysate ID according to positive signal site, according to lysate database information, find corresponding protein name, NCBI typing number (accessionnumber), protein I D, the information such as albumen size.
As shown in Figure 7, monoclonal antibody Umab83 can identify CD56 albumen specifically to result on OriGene protein chip, shows that the specificity of monoclonal antibody Umab83 is better.
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 (10)
1. a monoclonal antibody, is characterized in that, it is combined with CD56 protein-specific.
2. monoclonal antibody according to claim 1, is characterized in that, the hybridoma cell strain being CGMCCNo.11081 by deposit number produces.
3. the application of monoclonal antibody according to claim 1 and 2 in the immunodetection instrument for the preparation of detection CD56 albumen.
4. application according to claim 3, is characterized in that, described immunodetection instrument is reagent, test kit, chip or test paper.
5. an immunologic combined detection reagent kit, is characterized in that, comprises the monoclonal antibody described in claim 1 or 2.
6. monoclonal antibody as claimed in claim 1 or 2 is for the preparation of the application in the reagent of tagged tissue cell or test kit.
7. application according to claim 6, is characterized in that, described histocyte is natural killer cell, NK sample T cell, nerve, neuroendocrine tissue or related neoplasms.
8. a test kit for tagged tissue cell, is characterized in that, comprises the monoclonal antibody described in claim 1 or 2.
9. test kit according to claim 8, is characterized in that, described histocyte is natural killer cell, NK sample T cell, nerve, neuroendocrine tissue or related neoplasms.
10. a hybridoma cell strain, is characterized in that, its deposit number is CGMCCNo.11081.
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| CN107236025A (en) * | 2017-03-30 | 2017-10-10 | 中山大学 | The polypeptide combined with CD56 molecular specificities and its application |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106749668A (en) * | 2016-12-13 | 2017-05-31 | 无锡傲锐东源生物科技有限公司 | Anti- IDO1 protein monoclonal antibodies and application thereof |
| CN107236025A (en) * | 2017-03-30 | 2017-10-10 | 中山大学 | The polypeptide combined with CD56 molecular specificities and its application |
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| CN107488231B (en) * | 2017-09-15 | 2020-10-30 | 四川大学 | anti-CD 56 antibodies and uses thereof |
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Application publication date: 20160120 |