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

A hybridoma cell line, potato technology, applied in the direction of antiviral immunoglobulin, analytical materials, instruments, etc.

Active Publication Date: 2020-07-17
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 its application
  • Hybridoma cell line secreting anti-potato virus m monoclonal antibody and its application
  • Hybridoma cell line secreting anti-potato virus m monoclonal antibody and its application

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Embodiment Construction

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

[0019] A kind of described hybridoma cell secretes anti-potato M virus monoclonal antibody, and anti-potato M virus monoclonal antibody ascitic fluid indirect ELISA titer reaches 10- 7 , the antibody type and subclass are IgG1, kappa light chain, and have specific immune reaction with the 34kDa coat protein of potato virus M. Using ACP-ELISA and dot-ELISA methods, it was found that the monoclonal antibody detected the crude extraction of infected PVM potato leaf tissue. The sensitivity of the solution reaches 1:163840 and 1:10240 dilution (w / v, g / mL).

[0020] Anti-potato virus M monoclonal anti...

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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 invention relates to the field of biotechnology, in particular to a hybridoma cell line secreting anti-potato M virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Potato virus M (PVM) was first reported in the United States in 1923, which can cause 15% to 45% loss of potato yield. PVM is a member of the genus Carlavirus of the family Betaflexiviridae, and the virus particles are curved filaments of 610-700nm×12-15nm. Its genome is a single-stranded positive-sense RNA molecule of about 8500bp, with a cap structure at its 5' end, a poly(A) tail at its 3' end, and six open reading frames (ORFs). ORF 1 encodes a polypeptide containing methyltransferase, helicase and polymerase domains and is important for PVM RNA replication. Overlapping ORFs 2, 3, and 4 encode a triple-gene linkage (TGB)-encoded protein associated with viral movement. ORF5 and ORF 6 encode a 34 kDa viral capsid protein (CP) and a...

Claims

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Application Information

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Patent Type & Authority Patents(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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