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Hybridoma cell line secreting anti-rabies virus m protein monoclonal antibody and its application

A hybridoma cell line, monoclonal antibody technology, applied in anti-viral immunoglobulin, analytical materials, instruments, etc., can solve the problem that the fine epitope recognized by M protein monoclonal antibody is unclear and restricts anti-M protein monoclonal antibody. application, lack of biological characteristics of monoclonal antibodies, etc., to achieve the effect of excellent biological characteristics

Active Publication Date: 2020-10-13
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 line secreting anti-rabies virus m protein monoclonal antibody and its application
  • Hybridoma cell line secreting anti-rabies virus m protein monoclonal antibody and its application
  • Hybridoma cell line secreting anti-rabies virus m protein monoclonal antibody and its application

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Embodiment 1

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

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

[0025] According to the CVS-11 strain registered in GenBank (Liu Juan, et al. BECN1-dependent CASP2 incomplete autophagy induction by binding to rabies virus phosphoprotein. Autophagy, 2017, 13(4): 739-753) M protein gene (serial number: ADJ29910. 1) Design specific primers based on the nucleotide sequence, and use the extracted viral RNA reverse transcription product as a template to amplify the M protein gene, then clone the M protein gene into the pET-28a(+) vector, and verify that it is correct by sequencing Competent cells were transformed into E.coli BL21(DE3), induced with IPTG (1 mM) at 37°C for about 5 hours, and 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 electr...

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Abstract

The invention discloses a hybridoma cell strain which secretes a monoclonal antibody against rabies virus M protein and an application thereof, belonging to the field of biotechnology. The hybridoma cell line is classified and named as hybridoma cell line 4A1, which was deposited in the China Type Culture Collection on April 1, 2019, at the address: Wuhan, Wuhan, China, and the deposit number is CCTCC NO: C201947. The monoclonal antibody prepared by the hybridoma cell line has high titer, good specificity and excellent biological characteristics; it has identified the fine antigen variant epitope that recognizes the RABV M protein and can be used to distinguish the vaccine Flury strain from other RABV strains ; It is applied to the preparation of a kit for detecting rabies virus RABV, which can detect RABV infection and differential diagnosis vaccine Flury 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 Patents(China)
IPC IPC(8): C07K16/10C12N5/20G01N33/577G01N33/569
CPCC07K16/10G01N33/577G01N33/56983C07K2317/34G01N2333/145
Inventor 周继勇
Owner ZHEJIANG UNIV
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