Hybridoma cell line of monoclonal antibody against African swine fever virus and secreted monoclonal antibody thereof

A hybridoma cell line and African swine fever virus technology, applied in the fields of biotechnology and cell engineering, can solve the problems of low immunogenicity and achieve high practical value.

Inactive Publication Date: 2010-09-15
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0006] Most strains of African swine fever virus are highly virulent, but their immunogenicity is low, and only a few proteins are immunogenic

Method used

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  • Hybridoma cell line of monoclonal antibody against African swine fever virus and secreted monoclonal antibody thereof
  • Hybridoma cell line of monoclonal antibody against African swine fever virus and secreted monoclonal antibody thereof
  • Hybridoma cell line of monoclonal antibody against African swine fever virus and secreted monoclonal antibody thereof

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Experimental program
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Embodiment Construction

[0015] The hybridoma cell line of the monoclonal antibody against African swine fever virus is preserved in CGMCC No. 3771 of the General Microbiology Center of the China Microbiological Culture Collection Management Committee, the preservation date: 2010-04-22, and the classification name: hybridoma cell line.

[0016] Below in conjunction with specific embodiment the present invention is described in further detail:

[0017] Technical scheme of the present invention is as follows:

[0018] 1. Establishment of cell lines

[0019] 1. Antigen Preparation

[0020] a) Expression of African swine fever P30 protein

[0021] According to the African swine fever (ASFV) P30 gene sequence in GenBank, a pair of primers were designed and synthesized, and the P30 gene fragment was amplified from the ASFV DNA by PCR method, which was cloned into the expression vector pET28b to construct the recombinant plasmid pET-P30 After sequencing verification, it was transformed into the expression...

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Abstract

The invention discloses a hybridoma cell line of a monoclonal antibody against African swine fever virus and the secreted monoclonal antibody thereof. The preparation method of the invention comprises the following steps: preparing a recombined P30 soluble antigen by prokaryotic expression; immunizing a BALB/c mouse; and finally fusing, screening and cloning by a hybridoma technology to obtain the hybridoma cell line which can stably secrete the monoclonal antibody against African swine fever virus P30 protein. The invention further discloses a method for preparing the monoclonal antibody with the cell line, an antibody purification method and a labeling method for horseradish peroxidase of the antibody. The monoclonal antibody can be used in detecting the African swine fever viral antibody in pig serum.

Description

technical field [0001] The invention belongs to the field of biotechnology and cell engineering, and relates to a hybridoma cell line secreting an anti-ASFV monoclonal antibody and the monoclonal antibody secreted thereto, as well as the purification and horseradish peroxidase labeling of the monoclonal antibody, which can be used Applied to the detection of African swine fever virus antibody in pig serum. Background technique [0002] African swine fever (ASF) is an acute, febrile, highly contagious infectious disease of pigs caused by African swine fever virus (ASFV). It is characterized by a short course of disease and a high fatality rate, which can be as high as 100%. The clinical symptoms and pathological changes are similar to acute swine fever, and it is easy to be misdiagnosed during diagnosis. (William, Hess Adv.African swine fever: a reassessment[J].Vet Sci Comp Med, 1981, 25:39~69) The World Organization for Animals (OIE) listed it as a class A disease, and our ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/08C07K1/13C12R1/91
Inventor 董志珍侯艳梅肖妍赵祥平王涛张瑞杨新梅
Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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