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Monoclonal antibody hybridoma cell line 1H6 for resisting infectious bursal disease virus VP2 protein

A technology of hybridoma cell line and monoclonal antibody, which is applied in the direction of antiviral immunoglobulin, virus/bacteriophage, virus, etc., can solve the problems of limited research and development, and achieve good competitiveness and specificity

Active Publication Date: 2019-10-25
JIANGSU ACAD OF AGRI SCI
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Problems solved by technology

[0010] The present invention aims at the problem that none of the monoclonal antibodies obtained by lymphocyte hybridoma technology can react with prokaryotically expressed rVP2, which limits the development of IBDV competition ELISA antibody detection kits. As a double screening antigen, to develop a monoclonal antibody against IBDV rVP2, as a competitive antibody for IBDV antibodies in chicken serum samples, and to provide materials for the development of IBDV antibody competition ELISA detection kits

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  • Monoclonal antibody hybridoma cell line 1H6 for resisting infectious bursal disease virus VP2 protein

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Embodiment Construction

[0024] In the early stage, our laboratory used purified IBDV as a screening antigen and obtained multiple monoclonal antibodies against IBDV through lymphocyte hybridoma technology. However, none of these monoclonal antibodies could react with prokaryotically expressed rVP2, which limited the competition of IBDV for ELISA antibody detection. Development of kits. The present invention is aimed at the problem of above monoclonal antibody, uses the recombinant VP2 protein of prokaryotic expression and the IBDVQL strain of purification as double screening antigen, develops the monoclonal antibody against IBDV rVP2, is used as the competing antibody of IBDV antibody in the chicken serum sample, for the development of IBDV Antibody competition ELISA detection kit provides materials.

[0025] The method is as follows: use the IBDV QL virulent strain isolated from the bursa of Fabricius tissue of chickens with failed immunization recently as an antigen, immunize BALB / c mice, and use t...

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Abstract

The invention relates to a monoclonal antibody hybridoma cell line 1H6 for resisting infectious bursal disease virus VP2 protein, and belongs to the technical field of biology. In the invention, BALB / c mice are immunized with purified IBDV QL strain antigen, splenocytes are prepared and fused to an SP2 / 0 myeloma cell line, double screening is carried out by IBDV recombinant VP2 (rVP2) protein expressed by prokaryote and purified IBDV QL strain to obtain the monoclonal antibody hybridoma cell line 1H6 which not only can react with rVP2 protein expressed by prokaryote, but also can react with IBDV, the antibody subclass is IgG1 kappa, and the antibody titers of induced ascites are 108 respectively. Sandwich ELISA assay shows that 1H6 has no cross-reaction with other four avian viruses; indirect immunofluorescence test proves that 1H6 has good specific reaction; and immunoblot analysis shows that 1H6 can produce specific protein bands with rVP2 protein and IBDV.

Description

[0001] 1. Technical field [0002] The invention relates to a monoclonal antibody hybridoma cell line 1H6 against infectious bursal virus VP2 protein, belongs to the field of biotechnology, and relates to antibody engineering technology. [0003] 2. Background technology [0004] Infectious bursal disease (Infectious Bursal Disease, IBD) is one of the main infectious diseases that cause serious economic losses to the poultry industry in my country and the world. Organs - Infectious diseases characterized by the bursa of Fabricius. In recent years, the emergence of a super-virulent strain (vvIBDV) has greatly increased the immune failure and mortality of chicks, resulting in huge indirect economic losses. [0005] Vaccination is the most effective way to prevent the disease. In order to evaluate the protective effect of the vaccine, it is necessary to analyze the antibody level of chickens before and after vaccination. At present, the main methods for evaluating chicken IBDV an...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/10G01N33/68G01N33/577C12R1/91
CPCC07K14/005C07K16/10G01N33/68G01N33/577C12N2720/10022
Inventor 欧阳伟王永山夏兴霞王晓丽钱晶王晶宇诸玉梅马孙婷
Owner JIANGSU ACAD OF AGRI SCI
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