CN104962524A - Hybridoma cell strain 2C1 capable of secreting CAV-2 monoclonal antibodies, monoclonal antibodies secreted by cell strain and application - Google Patents
Hybridoma cell strain 2C1 capable of secreting CAV-2 monoclonal antibodies, monoclonal antibodies secreted by cell strain and application Download PDFInfo
- Publication number
- CN104962524A CN104962524A CN201510333160.2A CN201510333160A CN104962524A CN 104962524 A CN104962524 A CN 104962524A CN 201510333160 A CN201510333160 A CN 201510333160A CN 104962524 A CN104962524 A CN 104962524A
- Authority
- CN
- China
- Prior art keywords
- cav
- cell strain
- hybridoma cell
- monoclonal antibody
- monoclonal antibodies
- 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.)
- Pending
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a hybridoma cell strain 2C1 capable of secreting CAV-2 monoclonal antibodies, the monoclonal antibodies secreted by the cell strain and application. The preservation number of the hybridoma cell strain 2C1 is CGMCC No. 10878. An indirect immunofluorescence result shows that the monoclonal antibodies secreted by the hybridoma cell strain 2C1 can specifically recognize CAV-2 viruses and do not react with MDCK (madin-darby canine kidney) cells. A result of a superposition experiment of the hybridoma cell strain 2C1 and two monoclonal antibodies secreted by another hybridoma cell strain shows that the two monoclonal antibodies are used for different antigenic epitopes of the CAV-2 viruses. The monoclonal antibody prepared by the invention can react with CAV-2 well and can be used for identifying a clinic isolated strain. The 2C1 monoclonal antibody and the 7D7 monoclonal antibody which are prepared by the invention are used for recognizing two different antigen determinants of the CAV-2, so that the monoclonal antibodies can be used for detecting preparation of CAV-2 pathogenic colloidal gold and establishment of a sandwiched ELISA (enzyme-linked immuno sorbent assay) method.
Description
Technical field
The invention belongs to biological technical field, relate to the hybridoma cell strain of a strain secretion CAV-2 monoclonal antibody and the monoclonal antibody of being secreted by this cell strain and application thereof.
Background technology
Adenovirus (Canln adenovll.us, CAV) is one of virus studying the most detailed in the biological characteristicses such as morphological structure, gene replication mechanism up to now.Adenovirus capsid is made up of three major structural proteins: fine prominent, six adjacent body and pentons, it is infectious strong, and morphological structure is clear, and scientific research is of many uses.At present, the known serotype of hepatitis infectiosa canis virus is 1 type and 2 types, although have immunological cross provide protection between the two, they are all not identical in pathogenicity bo preferendum.These two kinds of genomic homologys of serotype adenovirus are between 57-70%.Hepatitis infectiosa canis virus infection is the viral infectious of dog, mainly causes the hepatitis of dog, respiratory disease and eyes illness etc., also can cause wolf, fox, Mammals encephalitis.In recent years, both at home and abroad alleged occurrence two kinds mutually about but the adenovirus differed from one another: viral 1 type (CAV-1) is the cause of disease of infectious canine hepatitis and epizootic fox encephalitis, and can ocular damage be caused, 2 types (CAV-2) then can cause the infectious laryngotracheitis of dog, pup pneumonia and bowel symptoms.CAV-2 infects pathological change and often shows respiratory tract, performance atelectasis, congested, and companion liver in various degree becomes, bronchial lymph nodes is congested, and histological examination visible moderate pneumonia pathology, bronchiolar epithelium, septal cell, concha epithelium have intranuclear inclusion.
Monoclonal antibody technique is swift and violent from developing over more than 30 year of coming out, and has been successfully used to the research in many fields such as molecular biology, immunology at present.In veterinary field, monoclonal antibody has played vital role for disease detection treatment.Colloidal gold immuno-chromatography test paper strip is quick, accurate, special and sensitive diagnostic method, only need a small amount of sample, and sample does not need process, just accurate judgement can be made to whether catching an illness in the very short time, be beneficial to cultivation unit early isolation, early treatment, meet the requirement of cultivation unit of basic unit for disease quick diagnosis.Also do not report for the monoclonal antibody of CAV-2 and the development of diagnosis colloidal gold strip thereof at present.
Summary of the invention
An object of the present invention is to provide the hybridoma cell strain 2C1 of a strain secretion CAV-2 monoclonal antibody.
The hybridoma cell strain 2C1 of one strain secretion CAV-2 monoclonal antibody, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is No. 3 China Microbiological institutes in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, deposit number is CGMCC No.10878, and preservation date is on May 27th, 2015.
Two of object of the present invention is to provide the anti-CAV-2 viral monoclonal antibodies that a strain above-mentioned hybridoma cell strain 2C1 secretes.
Three of object of the present invention is to provide the application of anti-CAV-2 viral monoclonal antibodies in preparation detection CAV-2 cause of disease reagent that above-mentioned hybridoma cell strain 2C1 secretes, and is in particular in following two aspects:
1, the anti-CAV-2 viral monoclonal antibodies secreted with hybridoma cell strain 7D7 is for detecting the preparation of the immunity colloidal gold test paper strip of CAV-2 cause of disease;
2, the anti-CAV-2 viral monoclonal antibodies secreted with hybridoma cell strain 7D7 is for detecting the foundation of CAV-2 cause of disease sandwich ELISA method.
Hybridoma cell strain 7D7, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is No. 3 China Microbiological institutes in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and deposit number is CGMCC No.10877, and preservation date is on May 27th, 2015.
The CAV-2 virus inoculation that laboratory is preserved by the present invention is bred on mdck cell, gathers in the crops virus, multigelation three times, the centrifugal 10min of ultrasonic 30min, 12000r/min, remove cell debris and larger impurity when pathology appears in the cell when 80%.Get supernatant, 4 DEG C, the centrifugal 2.5h of hypervelocity 28000r/min, get precipitation, resuspended with PBS, the centrifugal 10mln of 12000r/min, get its supernatant and cross discontinuous sucrose density gradient (60%-40%-30%), the centrifugal 2.5h of 28000r/min, gets its white portion, adds a large amount of PBS desugar, the centrifugal 2.5h of 28000r/min, get its precipitation, resuspended with 1.5ml PBS, the final virus obtaining CAV-2 purifying.
ELISA method detected result shows: the CAV-2 virus of purifying can with CAV-2 positive antibody generation specific reaction, show that it has good antigenicity.
Get virus and freund's adjuvant 1: 1 mixing and emulsifying of appropriate purifying, immunity BALB/C mice in 6 week age, get immune mouse spleen cell and SP2/0 cytogamy according to a conventional method, by the virus of purifying as detectable antigens, set up indirect EL1SA method and carry out the strain of detection screening positive cell clone to there being the cell conditioned medium liquid merging hole, finally obtain the hybridoma cell clone strain of the single secreting specificity antibody of two strains, respectively called after 2C1,7D7.
Indirect immunofluorescence result shows: the monoclonal anti physical efficiency specific recognition CAV-2 of 2C1 hybridoma cell strain secretion is viral and do not react with mdck cell.
2C1 prepared by the present invention shows with the superposition experimental result of two monoclonal antibodies that another hybridoma cell strain 7D7 secretes: these two monoclonal antibodies are the monoclonal antibody for the different epitope of CAV-2 virus.
Major advantage of the present invention is as follows:
1, the monoclonal anti physical efficiency prepared by the present invention well and CAV-2 reaction, can be used for the qualification of clinical isolated strain.
2, the 2C1 monoclonal antibody prepared by the present invention and 7D7 monoclonal antibody are the monoclonal antibodies of two different antigenic determinants identifying CAV-2, and therefore the foundation of monoclonal anti physical efficiency of the present invention for the preparation and sandwich ELISA method that detect CAV-2 cause of disease Radioactive colloidal gold is described.
Preservation information
One, the hybridoma cell strain 2C1 of anti-CAV-2 monoclonal antibody is secreted
Classification And Nomenclature: the hybridoma cell strain secreting anti-CAV-2 monoclonal antibody;
Depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center;
Preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City China Microbiological institute;
Deposit number: CGMCC No.10878;
Preservation date: on May 27th, 2015.
Two, the hybridoma cell strain 7D7 of anti-CAV-2 monoclonal antibody is secreted
Classification And Nomenclature: the hybridoma cell strain secreting anti-CAV-2 monoclonal antibody;
Depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center;
Preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City China Microbiological institute;
Deposit number: CGMCC No.10877;
Preservation date: on May 27th, 2015.
Accompanying drawing explanation
Fig. 1 is CAV-2 specificity object fragment, 1: molecular weight standard DL 1000; 2,3,4: different concns CAV-2 virus PCR product; 3: negative control;
Fig. 2 is the analysis of CAV-2 specificity object fragment PCR products sequencing result;
Fig. 3 is the qualification of CAV-2 virus protein activities after purifying;
Fig. 4 is the IIF qualification of anti-CAV-2 monoclonal antibody, a:2C1 monoclonal antibody, b:7D7 monoclonal antibody, c: positive mouse serum control, and d:sp2/0 supernatant contrasts.
Embodiment
Below in conjunction with accompanying drawing, technical scheme of the present invention is further described; but be not limited thereto; everyly technical solution of the present invention modified or equivalent to replace, and not departing from the spirit and scope of technical solution of the present invention, all should be encompassed in protection scope of the present invention.
The invention provides a kind of preparation method identifying the anti-CAV-2 monoclonal antibody of different epitope, particular content is as follows:
1 materials and methods
1.1 materials, laboratory animal and reagent
Experiment BALB/c mouse is provided by the Chinese Academy of Agricultural Sciences's Harbin veterinary institute Experimental Animal Center; Freund's adjuvant, selective reagents HAT, HT, fusogen PEG are purchased from Sigma company; Standard foetal calf serum, substratum RPM11640 are purchased from Gibco company; Microbiotic is purchased from the precious biotechnology company limited in Dalian.CAV-2 is preserved by veterinary institute disease diagnosis center, Harbin.
The cultivation of 1.2 CAV-2 and the extraction of genomic dna
By virus inoculation on mdck cell, cultivate with the DMEM nutrient solution containing 10% bovine serum, after 90% cell generation pathology, results virus, prepares genomic DNA template by the method that viral cultures is extracted on test kit specification sheets by viral DNA.
1.3 PCR qualifications
Utilize the design of DNSTAR biological software for the primer of CAV-2, upstream primer is: 5 '-CGCGCTGAACATTACTACCTTGTC-3 ', and downstream primer is: 5 '-CCTAGAGCACTTCGTGTCCGCTT-3 '.Take viral DNA as template, carry out pcr amplification by high-fidelity Taq enzyme (Pyrobest archaeal dna polymerase, MB1 company), PCR primer observes the position of band through agarose gel electrophoresis, PCR primer is sent order-checking simultaneously.
The purifying of 1.4 CAV-2
The virus inoculation being accredited as CAV-2 is bred on mdck cell, gathers in the crops virus, multigelation three times, ultrasonic 30min, 12000r/min centrifugal 10min when pathology appears in the cell when 80%, remove cell debris and larger impurity.Get supernatant, 4 DEG C, the centrifugal 2.5h of hypervelocity 28000r/min, get precipitation, resuspended with PBS, the centrifugal 10min of 12000r/min, gets its supernatant and crosses discontinuous sucrose density gradient (60%-40%-30%), the centrifugal 2.5h of 28000r/min, get its white portion, add a large amount of PBS desugar, the centrifugal 2.5h of 28000r/min, get its precipitation, resuspended with 1.5ml PBS.
The activation analysis of 1.5 purified viruses
Get the CAV-2 after purifying with the concentration bag in 2 μ g/ holes by 96 orifice plates, using the positive serum of the CAV-2 infected mice suitably diluted as primary antibodie, the sheep anti mouse 1gG of horseradish peroxidase-labeled two resists, and develops the color with tmb substrate.
The preparation of 1.5 monoclonal antibodies
Get virus and Freund's complete adjuvant 1: 1 mixing and emulsifying of appropriate purifying, inoculation 6-8 BALB/C mice in age in week, after this carried out two, three immunity at interval of two weeks with equivalent amount of antigen and Freund's incomplete adjuvant emulsification.Three exempt from 3 days after, get immune mouse spleen cell and SP2/0 cell with 4: 1-10: 1 ratio under the effect of PEG1500, carry out cytogamy, cultivate with HAT Selective agar medium after merging, within every three days, change a nutrient solution partly to change liquid method, within first 6 days, use HAT nutrient solution, then HAT nutrient solution is replaced with HT nutrient solution gradually partly to change liquid method, when clone cell covers with at the bottom of hole 1/10, by the virus of purifying as detectable antigens, carry out indirect EL1SA to there being the cell conditioned medium liquid merging hole and detect the strain of screening positive cell clone.The positive cell clone strain limiting dilution assay screened is carried out mono-clonal, detects by indirect EL1SA method, until positive rate reaches 100%, when obtaining the hybridoma cell clone of single secreting specificity antibody, enlarged culturing respectively, and frozen in liquid nitrogen.
1.6 indirect immunofluorescence assay
Mdck cell is infected with the CAV-2 of clone, when pathology reaches 80% with the paraformaldehyde fixed cell 15min of 4%, then respectively by the positive mouse serum of known CAV-2, CAV-2 monoclonal antibody and SP2/0 cells and supernatant as primary antibodie, correspondingly resist using the sheep anti mouse 1gG of F1TC mark as two, carry out indirect immunofluorescene assay according to a conventional method, establish the normal mdck cell control group of uninfecting virus simultaneously.
The hypotype qualification of 1.7 monoclonal antibodies
According to monoclonal antibody hypotype identification kit operation instructions, respectively hypotype qualification is carried out to the monoclonal antibody obtained above.
The superposition experiment of 1.8 monoclonal antibodies
By the virus of purifying as envelope antigen, apply indirect EL1SA method, respectively by after a strain monoclonal antibody effect 1h, another strain monoclonal antibody is remake with 1h, then the sheep anti mouse 1gG bis-adding horseradish mark resists, with TMB colour developing, establish the control group of hatching single monoclonal antibody, two monoclonal antibodies be significantly increased as OD value are the monoclonal antibody for different epitope simultaneously.
The preparation of 1.9 ascites and the mensuration of tiring
Method routinely carries out the preparation of odd contradictive hydroperitoneum.The mensuration of Mab supernatant and titer of ascites is carried out above by the ELISA method set up.
2 results
The qualification result of 2.1 CAV-2
With different concns CAV-2 strain genomic dna for template, adopt CAV-2 special primer to carry out pcr amplification, result amplifies the band of 1.00kb, conforms to, as shown in Figure 1 with expection object fragment.
Row that PCR primer checks order are uploaded GENEBANK and carries out blast sequence alignment, as shown in Figure 2, result display gained object fragment is the E3 protein gene sequence in CAV-2 genome, illustrates that identified virus is CAV-2 in interpretation of result.
2.2 CAV-2 activity identification
With the CAV-2 albumen bag after purifying by 96 orifice plates, using mouse positive serum and mouse negative serum as primary antibodie, be two anti-ly carry out the detection of EL1SA method with the sheep anti mouse 1gG of HRP mark, result positive serum hole is aobvious blue, illustrate that the albumen after purifying has antigen reactivity, as shown in Figure 3.
The acquisition of 2.3 hybridoma cell strains
Through cytogamy and indirect EL1SA method preliminary screening, at the beginning of result, be sieved to 15 strain positive colonies; Carry out again 3 mono-clonals, until the positive rate in all clone holes reaches 100%, finally obtain the energy of 2 strain stably excretings and the cell strain of CAV-2 albumen generation positive reaction, called after 2C1,7D7, distinguished frozen respectively.
The unique identification test of 2.4 monoclonal antibodies
2 strain monoclonal antibodies of preparation in experiment are carried out indirect immunofluorescene assay with the mdck cell that CAV-2 infects respectively, result shows the 2 strain monoclonal antibodies obtained and all has specific fluorescence to the cell that CAV-2 infects, present the remarkable positive, and be feminine gender with not connecing malicious mdck cell without specificity fluorescent, the contrast of CAV-2 positive serum and negative serum is set up, as shown in Figure 4.Show that prepared monoclonal antibody has good specificity, energy specific recognition CAV-2 virus, can be used for test in laboratory and identifies the strain be separated.
The qualification of 2.5 monoclonal antibody hypotypes
Carried out hypotype qualification according to monoclonal antibody hypotype identification kit operation instructions to the 2 strain monoclonal antibodies obtained, all monoclonal antibodies of result are IgG1 type, and the light chain of all monoclonal antibodies is k chain.
2.6 monoclonal antibodies are for the qualification of different epitope
Monoclonal antibody superposition experiment in, 2C1 and 7D7 monoclonal antibody superposition after OD value have obvious increase, 2C1 secretion monoclonal antibody identification be an epitope A, 7D7 secrete monoclonal antibody identification be another one epitope B.As shown in table 1.So, this experiment obtains the hybridoma cell strain of the different epitope monoclonal antibody of two strain secretion identification two, 2C1 and 7D7 two strain monoclonal antibody is the monoclonal antibody of two the different antigenic determinants identifying virus, can be used for the foundation of the double-antibodies sandwich ELISA of the preparation and determination methods CAV-2 cause of disease of the immunity colloidal gold test paper strip detecting CAV-2 cause of disease.
Superposition experiment between the different monoclonal antibody of table 1
--: after representing superposition, OD value does not significantly change; ++ after representing superposition, OD value has obvious rising
The preparation of 2.7 ascites and the mensuration of tiring
Adopt indirect ELISA method to measure hybridoma cell strain 2C1 and tiring of 7D7 ascites and be respectively 1: 104200 and 1: 204800, tire 1: 640 apparently higher than corresponding hybridoma supernatant.Titer of ascites prepared by explanation is high, can be used for the development detecting CAV-2 cause of disease immunity colloidal gold test paper strip.
Claims (6)
1. the hybridoma cell strain 2C1 of a strain secretion CAV-2 monoclonal antibody, deposit number is CGMCC No.10878, and preservation date is on May 27th, 2015.
2. the anti-CAV-2 monoclonal antibody of the different epitopes of identification of hybridoma cell strain 2C1 secretion described in claim 1.
3. the identification different epitope CAV-2 monoclonal antibody that hybridoma cell strain 2C1 according to claim 2 secretes is detecting the application in CAV-2 etiological diagnosis reagent.
4. identification according to claim 3 different epitope CAV-2 monoclonal antibody is detecting the application in CAV-2 etiological diagnosis reagent, it is characterized in that the identification difference epitope CAV-2 monoclonal antibody that described hybridoma cell strain 2C1 secretes and the anti-CAV-2 monoclonal antibody that hybridoma cell strain 7D7 secretes are for detecting the preparation of the immunity colloidal gold test paper strip of CAV-2 cause of disease.
5. identification according to claim 3 different epitope CAV-2 monoclonal antibody is detecting the application in CAV-2 etiological diagnosis reagent, it is characterized in that the identification difference epitope CAV-2 monoclonal antibody that described hybridoma cell strain 2C1 secretes and the anti-CAV-2 viral monoclonal antibodies that hybridoma cell strain 7D7 secretes are for detecting the foundation of CAV-2 cause of disease sandwich ELISA method.
6. the different epitope CAV-2 of the identification according to claim 4 or 5 monoclonal antibody is detecting the application in CAV-2 etiological diagnosis reagent, it is characterized in that described hybridoma cell strain 7D7 deposit number is CGMCC No.10877, preservation date is on May 27th, 2015.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510333160.2A CN104962524A (en) | 2015-06-16 | 2015-06-16 | Hybridoma cell strain 2C1 capable of secreting CAV-2 monoclonal antibodies, monoclonal antibodies secreted by cell strain and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510333160.2A CN104962524A (en) | 2015-06-16 | 2015-06-16 | Hybridoma cell strain 2C1 capable of secreting CAV-2 monoclonal antibodies, monoclonal antibodies secreted by cell strain and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104962524A true CN104962524A (en) | 2015-10-07 |
Family
ID=54216621
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510333160.2A Pending CN104962524A (en) | 2015-06-16 | 2015-06-16 | Hybridoma cell strain 2C1 capable of secreting CAV-2 monoclonal antibodies, monoclonal antibodies secreted by cell strain and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104962524A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105891491A (en) * | 2016-04-11 | 2016-08-24 | 洛阳普莱柯万泰生物技术有限公司 | Kit and application thereof |
| CN108872570A (en) * | 2018-09-20 | 2018-11-23 | 扬州大学 | Double sandwich ELISA kits of detection 4 type aviadenovirus of serum based on Fiber1 albumen |
| CN109957009A (en) * | 2019-03-28 | 2019-07-02 | 中国人民解放军军事科学院军事医学研究院 | Anti-human 7 type adenovirus antibody 2-1H and its application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4487829A (en) * | 1982-03-23 | 1984-12-11 | Massachusetts Institute Of Technology | Production and use of monoclonal antibodies against adenoviruses |
-
2015
- 2015-06-16 CN CN201510333160.2A patent/CN104962524A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4487829A (en) * | 1982-03-23 | 1984-12-11 | Massachusetts Institute Of Technology | Production and use of monoclonal antibodies against adenoviruses |
Non-Patent Citations (2)
| Title |
|---|
| C.A. WHETSTONE: "Monoclonal Antibodies to Canine Adenoviruses 1 and 2 that are Type-Specific by Virus Neutralization and Immunofluorescence", 《VETERINARY MICROBIOLOGY》 * |
| WHETSTONE,C.A: "Should the criteria for species distinction in adenoviruses be reconsidered?Evidence from canine adenoviruses 1 and 2", 《INTERVIROLOGY》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105891491A (en) * | 2016-04-11 | 2016-08-24 | 洛阳普莱柯万泰生物技术有限公司 | Kit and application thereof |
| CN108872570A (en) * | 2018-09-20 | 2018-11-23 | 扬州大学 | Double sandwich ELISA kits of detection 4 type aviadenovirus of serum based on Fiber1 albumen |
| CN109957009A (en) * | 2019-03-28 | 2019-07-02 | 中国人民解放军军事科学院军事医学研究院 | Anti-human 7 type adenovirus antibody 2-1H and its application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | A novel neutralizing monoclonal antibody targeting the N-terminal domain of the MERS-CoV spike protein | |
| Boldorini et al. | Serological evidence of vertical transmission of JC and BK polyomaviruses in humans | |
| Meyer et al. | Serologic assessment of possibility for MERS-CoV infection in equids | |
| Origgi et al. | Enzyme-linked immunosorbent assay for detecting herpesvirus exposure in Mediterranean tortoises (spur-thighed tortoise [Testudo graeca] and Hermann's tortoise [Testudo hermanni]) | |
| CN107937349A (en) | Stablize cell line and its preparation and application of expression African swine fever virus P54 albumen | |
| CN105018431A (en) | Hybridoma cell line 7D7 secreting CAV-2 monoclonal antibody, monoclonal antibody secreted by hybridoma cell line 7D7, and applications of monoclonal antibody | |
| CN103163299A (en) | Avian leukosis double-antibody sandwich enzyme-linked immuno sorbent assay (ELISA) antigen detection kit | |
| CN104962524A (en) | Hybridoma cell strain 2C1 capable of secreting CAV-2 monoclonal antibodies, monoclonal antibodies secreted by cell strain and application | |
| Yang et al. | Single-domain antibodies as promising experimental tools in imaging and isolation of porcine epidemic diarrhea virus | |
| CN104220090A (en) | Enterovirus 71 specific antibodies and uses thereof | |
| Zhou et al. | Detection of ALV p27 in cloacal swabs and virus isolation medium by sELISA | |
| Scheffer et al. | Rabies virus in naturally infected bats in the State of São Paulo, Southeastern Brazil | |
| CN104498439A (en) | Hybridoma cell, PCV2 monoclonal antibody and application thereof | |
| CN113604438B (en) | Monoclonal antibody for resisting tilapia lake virus, cell strain and application thereof | |
| Janowski et al. | High seropositivity rate of neutralizing antibodies to astrovirus VA1 in human populations | |
| Dubuisson* et al. | Proteins specified by bovine herpesvirus type 4: structural proteins of the virion and identification of two major glycoproteins by using monoclonal antibodies | |
| CN103923881B (en) | Hepatitis A virus (HAV) monoclonal antibody and application thereof | |
| CN103602636B (en) | The monoclonal antibody of anti-sheep of virus B2L albumen and application thereof | |
| CN106701687A (en) | Hybridoma cell strain and rabies virus phosphoprotein monoclonal antibody generated by same | |
| CN101591390B (en) | H5N1 derived avian influenza virus NP resistant monoclonal antibody and application thereof | |
| Li et al. | Development of a sandwich ELISA for the detection of Chinese sacbrood virus infection | |
| Timoshicheva et al. | Use of hexon as an antigen for the production of monoclonal antibodies capable of detecting multiple adenovirus types | |
| CN106749647A (en) | Salmon trout IHNV monoclonal antibodies and detection kit | |
| CN106198974A (en) | The ELISA detection kit of IHNV antibody in a kind of quick detection rainbow trout serum | |
| CN116790509B (en) | Monoclonal hybridoma cell strain secreting anti-porcine pseudorabies virus gB protein 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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20151007 |