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Hybridoma cell line secreting anti-potato virus M monoclonal antibody and application of monoclonal antibody

A hybridoma cell line, potato technology, applied in antiviral immunoglobulin, analytical materials, biochemical equipment and methods, etc.

Active Publication Date: 2019-09-20
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the establishment of serological methods depends on specific high-quality virus antibodies

Method used

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  • Hybridoma cell line secreting anti-potato virus M monoclonal antibody and application of monoclonal antibody
  • Hybridoma cell line secreting anti-potato virus M monoclonal antibody and application of monoclonal antibody
  • Hybridoma cell line secreting anti-potato virus M monoclonal antibody and application of monoclonal antibody

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Embodiment Construction

[0018] The hybridoma cell line 1E1 that secretes anti-potato M virus monoclonal antibody was deposited in the General Microbiology Center of the Chinese Microbial Culture Collection Management Committee of the Institute of Microbiology, Chinese Academy of Sciences on January 25, 2019. The deposit number is CGMCC No.17284, which can secrete anti-potato M virus monoclonal antibody.

[0019] A said hybridoma cell secretes anti-potato M virus monoclonal antibody, and the anti-potato M virus monoclonal antibody ascites indirect ELISA titer reaches 10- 7 The antibody type and subclass are IgG1 and kappa light chain, which have a specific immune reaction with the 34kDa coat protein of potato M virus. Analysis using ACP-ELISA and dot-ELISA methods found that the monoclonal antibody was used to detect the crude extract of the diseased leaf tissue of the potato infected with PVM. The sensitivity of the solution reaches 1:163840 and 1:10240 dilution (w / v, g / mL).

[0020] The anti-potato M vir...

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Abstract

The invention discloses a hybridoma cell line secreting an anti-potato virus M monoclonal antibody and an application of the monoclonal antibody. BALB / c mice are immunized with purified PVM virus particles as antigens, and the hybridoma cell line 1E1 secreting the anti-PVM monoclonal antibody is obtained through cell fusion, screening and cloning and has the preservation number of CGMCC No.17284. The ascites indirect ELISA titer of the monoclonal antibody secreted by the cell line is 10<-7>. The antibody type and subclass are IgG1 and a kappa light chain. The monoclonal antibody has specific immune reaction to the PVM, but does not have reaction with potato virus S, potato virus A, potato virus X, potato virus Y and potato leaf roll virus. The ACP-ELISA, dot-ELISA and Tissueprint-ELISA detection methods for detecting the PVM in potato plants are established by using the 1E1 monoclonal antibody, wherein the sensitivities of the ACP-ELISA and dot-ELISA methods for detecting diseased leaves is 1:163840 and 1:10240 times dilution (w / v, g / mL) respectively.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting monoclonal antibodies against potato M virus and the application of the monoclonal antibodies thereof. Background technique [0002] Potato virus M (PVM) was first reported in the United States in 1923, which can cause a 15% to 45% loss of potato yield. PVM belongs to the Betaflexiviridae family (Betaflexiviridae), a member of the Carlavirus genus, and the virus particles are curved filaments of 610-700nm×12-15nm. Its genome is a single-stranded sense RNA molecule of about 8500 bp with a cap structure at the 5'end, a poly(A) tail at the 3'end, and 6 open reading frames (ORF). ORF 1 encodes a polypeptide containing methyltransferase, helicase, and polymerase domains, which are important for PVM RNA replication. Overlapping ORF 2, 3, and 4 code for a three-gene linkage structure (TGB)-encoding protein associated with virus movement. ORF5 and ORF ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/10G01N33/569G01N33/577C12R1/91
CPCC07K16/10C07K2317/33C07K2317/35G01N33/56983G01N33/577
Inventor 吴建祥张彧周雪平钱亚娟
Owner ZHEJIANG UNIV
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