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Hybridoma cell strain to secrete anti-Rabies virus M protein monoclonal antibody and application thereof

A technology of hybridoma cell lines and monoclonal antibodies, applied in antiviral immunoglobulins, analytical materials, biochemical equipment and methods, etc., can solve the problem of unclear fine epitopes recognized by M protein monoclonal antibodies and lack of monoclonal Biological characteristics of antibodies, restrictions on the application of anti-M protein monoclonal antibodies, etc., to achieve the effect of excellent biological characteristics

Active Publication Date: 2019-07-05
ZHEJIANG UNIV
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AI Technical Summary

Problems solved by technology

Monoclonal antibodies to RABVM protein are rarely reported, and the fine epitopes recognized by M protein monoclonal antibodies are still unclear, and the lack of understanding of the biological characteristics of these monoclonal antibodies also limits the use of anti-M protein monoclonal antibodies in virus diagnosis. application in , and studies on the structure and antigenic properties of M protein

Method used

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  • Hybridoma cell strain to secrete anti-Rabies virus M protein monoclonal antibody and application thereof
  • Hybridoma cell strain to secrete anti-Rabies virus M protein monoclonal antibody and application thereof
  • Hybridoma cell strain to secrete anti-Rabies virus M protein monoclonal antibody and application thereof

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Embodiment 1

[0023] (1) Establishment of monoclonal antibody-secreting hybridoma cell lines

[0024] 1. Prokaryotic expression and purification of CVS-11M protein

[0025] According to the CVS-11 strain registered in GenBank (Liu Juan, et al.BECN1-dependentCASP2incomplete autophagy induction by binding to rabies virusphosphoprotein.Autophagy,2017,13(4):739-753) M protein gene (sequence number: ADJ29910.1 ) nucleotide sequence to design specific primers, and use the extracted viral RNA reverse transcription product as a template to amplify the M protein gene, and then clone the M protein gene into the pET-28a(+) vector, after the sequence verification is correct Competent cells E.coli BL21(DE3) were transformed, and after induction with IPTG (1 mM) for about 5 h at 37°C, the recombinant M protein was purified with a Ni-NTA column (purchased from QiAGEN). The expression and purification of recombinant M protein were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-...

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Abstract

The invention discloses a hybridoma cell strain to secrete anti-Rabies virus M protein monoclonal antibody and application thereof and belongs to the technical field of biology. The hybridoma cell strain has a category name of hybridoma cell strain 4A1, collected in China Center for Type Culture Collection, Wuhan University, Wuhan, China under CCTCC NO: C201947. A monoclonal antibody prepared withthe hybridoma cell strain has high titer, good specificity and good biological features; fine antigen variant epitope of the monoclonal antibody in recognizing RABV M protein is identified, so that the monoclonal antibody is suitable for distinguishing vaccine Flury virus strains and other RABV strains; when applied to the preparation of kits to detect RABV (Rabies virus), the monoclonal antibodyis capable of detecting RABV infections and identifying and diagnosing vaccine Flury virus strain and other RABV strains.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting anti-rabies virus M protein monoclonal antibody and its application. Background technique [0002] Rabies is a highly lethal zoonotic disease caused by rabies virus (RABV). After the onset of RABV infection, the fatality rate is almost 100%, which is the acute infectious disease with the highest mortality rate in human history so far. Currently there is no treatment, and RABV infection can only be prevented through vaccination. Rabies cases have been reported in more than 150 countries around the world, and about 50,000 to 70,000 people die from rabies every year, 95% of which occur in Africa and Asia. China's human rabies infection cases are second only to India, ranking second in Asia. Dogs are the most common viral reservoir of RABV, and 99% of human rabies infections are mediated by canine transmission. [0003] RABV is a member of Rhabdoviri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N5/20G01N33/577G01N33/569
CPCC07K16/10G01N33/56983G01N33/577
Inventor 周继勇
Owner ZHEJIANG UNIV
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