Recombinant swine fever virus E2 protein swine source monoclonal antibody and preparation method and application thereof

A monoclonal antibody, swine fever virus technology, applied in antiviral immunoglobulins, biochemical equipment and methods, viruses/phages, etc., can solve problems such as instability, hybridoma cells cannot be stored for a long time, and antibody cell viability declines.

Active Publication Date: 2018-05-01
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0005] In the traditional method of preparing monoclonal antibodies, it takes a long time to prepare antibodies, and because hybridoma cells cannot be stored for a long time, it may cause the decline of antibody cell viability and loss of resistance.
In order to overcome the problems of unstable hybridoma cells producing monoclonal antibodies and easy loss of r

Method used

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  • Recombinant swine fever virus E2 protein swine source monoclonal antibody and preparation method and application thereof
  • Recombinant swine fever virus E2 protein swine source monoclonal antibody and preparation method and application thereof
  • Recombinant swine fever virus E2 protein swine source monoclonal antibody and preparation method and application thereof

Examples

Experimental program
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Embodiment 1

[0042] Example 1 Preparation of recombinant swine fever virus E2 protein porcine monoclonal antibody

[0043] 1. Materials and methods

[0044] 1.1 Cells, viruses and plasmids

[0045]Human embryonic kidney cells (HEK293T) and pig kidney cells (PK-15) were purchased from the American Type Culture Collection; Chinese hamster ovary cells (CHO) were purchased from the Cell Bank of the Chinese Academy of Sciences; suspension HEK293 cells were purchased from Harbin National Engineering Research Institute for Animal Biological Products Provided by Center Ltd. CSFV Shimen strain is preserved by our laboratory. Anti-CSFV E2 protein monoclonal antibody WH303 was donated by Professor Trevor W.Drew from the National Veterinary Laboratory of the United Kingdom. BVDV porcine-derived positive serum was donated by Professor Paul Becher of the OIE / EU Reference Laboratory for Swine Fever at Hannover Veterinary University.

[0046] HEK293T and PK-15 cells were cultured with DMEM containing ...

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Abstract

The invention discloses a recombinant swine fever virus E2 protein swine source monoclonal antibody and a preparation method and application thereof. Firstly, the recombinant swine fever virus E2 protein swine source monoclonal antibody is disclosed, an amino acid sequence of a heavy chain is shown in SEQ ID NO.1, and an amino acid sequence of a light chain is shown in SEQ ID NO.2. The invention also discloses a suspension HEK293 cell line which can stably express the recombinant swine fever virus E2 protein swine source monoclonal antibody. The recombinant swine fever virus E2 protein swine source monoclonal antibody has the advantages that the coding genes of the heavy chain and light chain are cloned into an eukaryotic expression vector, and the stable and high-efficiency expression ofthe recombinant swine fever virus E2 protein swine source monoclonal antibody is realized by the suspension HEK293 cell line; the reactogenicity and neutralizing activity are good, and the important development value is realized in development of novel swine fever virus diagnosing and treating preparations.

Description

technical field [0001] The present invention relates to a porcine monoclonal antibody of swine fever virus E2 protein, further relates to the preparation method of said porcine monoclonal antibody, and the present invention also relates to the preparation of porcine monoclonal antibody of swine fever virus E2 protein The invention relates to application in diagnostic reagents or antiviral preparations of classical swine fever virus, and belongs to the field of preparation of swine-derived monoclonal antibodies of recombinant classical swine fever virus E2 protein. Background technique [0002] Classical swine fever (CSF) is a severe infectious disease that seriously harms the pig industry and often causes huge economic losses. The disease is listed as an animal disease that must be declared by the World Organization for Animal Health (OIE). The pathogen of CSF is classical swine fever virus (CSFV). CSFV is an enveloped single-stranded positive-sense RNA virus with a genome l...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N15/13C12N5/10C12N15/867A61K39/42A61P31/14G01N33/68G01N33/577G01N33/569
CPCC07K16/1081C07K2317/14C07K2317/24C07K2317/33C07K2317/76C07K2317/92C12N15/86C12N2740/15043G01N33/56983G01N33/577G01N33/6854G01N2469/10
Inventor 仇华吉陈姝承李素罗玉子孙元
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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