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Cell strain of monoclonal antibodies of E2 protein resisting hog cholera virus and application thereof

A technology of monoclonal antibody and classical swine fever virus, which is applied in the direction of antiviral immunoglobulin, immunoglobulin, biochemical equipment and methods, and can solve problems such as clinical diagnosis and prevention and treatment difficulties

Active Publication Date: 2017-08-18
CHINA INST OF VETERINARY DRUG CONTROL
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AI Technical Summary

Problems solved by technology

At the same time, the mixed infection of multiple pathogens has also become a very serious phenomenon, which brings great difficulties to clinical diagnosis and prevention.

Method used

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  • Cell strain of monoclonal antibodies of E2 protein resisting hog cholera virus and application thereof
  • Cell strain of monoclonal antibodies of E2 protein resisting hog cholera virus and application thereof
  • Cell strain of monoclonal antibodies of E2 protein resisting hog cholera virus and application thereof

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Embodiment Construction

[0013] 1. Preparation of monoclonal antibody cell line against classical swine fever virus E2 protein

[0014] (1) Animal immunization Take five 6-week-old BALB / C female mice, mix and emulsify the eukaryotically expressed and purified classical classical swine fever virus E2 protein with an equal amount of Freund's complete adjuvant, and then put it at a dose of 100 μg / mouse Mice were injected at multiple points on the back; 14 days later, the E2 protein was mixed and emulsified with an equal amount of Freund's incomplete adjuvant, and the booster immunization was carried out according to the above method; 14 days later, the immunization was repeated once. After 14 days, blood was collected from the orbit, the serum was separated, and the antibody titer of the immunized mice was determined by indirect ELISA. Select the mouse with the highest antibody titer, and inject 100 μg E2 protein intraperitoneally. After 3 days, the spleen was taken for cell fusion.

[0015] (2) Cell f...

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Abstract

The invention relates to a cell strain of monoclonal antibodies of E2 protein resisting hog cholera virus and an application thereof. A hybridoma cell strain stably secreting the protein E2 resisting the hog cholera virus is obtained by screening, the monoclonal antibodies CSFV-1C8 secreted by the cell strain can generate specific reaction with the E2 protein of CSFV, and does not react with E2 protein of bovine viral diarrhoea / mucosal disease virus; and the monoclonal antibodies can recognize the E2 protein of the hog cholera virus specifically, and the epitope of recognition is FDFDGPDGL. The monoclonal antibodies and the epitope of the E2 protein of the hog cholera virus recognized by the monoclonal antibodies can be prepared into a reagent for detecting the CSFV, and thus laying foundation for establishing a serological detection method of antibodies of the hog cholera virus. A competitive ELISA kit for detecting the antibodies for the hog cholera and established by the invention has the advantages of specificity, sensitivity, simplicity and convenience in operation and the like and is suitable for large-scale screening of the antibodies for the hog cholera.

Description

technical field [0001] The invention relates to a monoclonal antibody cell strain of classical swine fever virus E2 protein and its application, belonging to the field of veterinary biological products. Background technique [0002] Classical Swine Fever (CSF) is a highly contagious disease caused by Classical Swine Fever Virus (CSFV), an important member of the Flaviviridae family Pestivirus. It is one of the epidemic diseases that must be reported according to the regulations of the World Health Organization (OIE), and it belongs to the first class of animal infectious diseases in my country. CSFV is an enveloped single-stranded positive-sense RNA virus with a diameter of 25-50 nm. In the same genus as CSFV are Bovine viral diarrhea virus (BVDV) and sheep border disease virus (Borderdisease virus, BDV). These three viruses have similar structures, cross-reactivity in serology, and similar host range. Pigs (both domestic and wild boar) are the only susceptible hosts. CSF...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/10G01N33/577G01N33/569C12R1/91
CPCC07K16/1081G01N33/56983G01N33/577G01N2469/20
Inventor 王琴徐璐张乾义赵启祖范学政邹兴启朱元源李翠
Owner CHINA INST OF VETERINARY DRUG CONTROL
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