Hybridoma cell strain secreting PPV (plum pox virus)-resistant monoclonal antibody and application of monoclonal antibody

A hybridoma cell line and plum pox virus technology, applied in the biological field, can solve the problems of poor detection sensitivity and low accuracy.

Active Publication Date: 2019-06-18
ZHEJIANG UNIV
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Problems solved by technology

At present, there have been reports on the preparation of PPV antibodies and serological methods, but the detectio

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  • Hybridoma cell strain secreting PPV (plum pox virus)-resistant monoclonal antibody and application of monoclonal antibody
  • Hybridoma cell strain secreting PPV (plum pox virus)-resistant monoclonal antibody and application of monoclonal antibody
  • Hybridoma cell strain secreting PPV (plum pox virus)-resistant monoclonal antibody and application of monoclonal antibody

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Embodiment Construction

[0018] The hybridoma cell line 4A8 secreting anti-plum pox virus monoclonal antibody was deposited in the General Microbiology Center of China Microbiological Culture Collection Management Committee, Institute of Microbiology, Chinese Academy of Sciences on January 25, 2019, and the preservation number is CGMCC No.17283. Monoclonal antibody against plum pox virus Chinese isolate.

[0019] A kind of described hybridoma cell secretes anti-plum pox virus monoclonal antibody, and anti-plum pox virus monoclonal antibody ascites indirect ELISA titer reaches 10 -7 , the antibody type and subclass are IgG1, kappa light chain, and have a specific immune reaction with the 36kDa coat protein of plum pox virus. Using TAS-ELISA and dot-ELISA methods, it was found that the monoclonal antibody detected the crude extract of PPV-infected leaves. Sensitivity reaches 1:20480 and 1:5120 dilutions.

[0020] The anti-plum pox virus monoclonal antibody can specifically react with plum pox virus, bu...

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Abstract

The invention discloses a hybridoma cell strain secreting a PPV (plum pox virus)-resistant monoclonal antibody and an application of the monoclonal antibody. Prokaryotic-expression PPV coat protein istaken as an antigen to immunize a BALB/c mouse, one hybridoma cell strain 4A8 capable of secreting the PPV-resistant monoclonal antibody is obtained by cell fusion, screening and cloning, and the preservation number of the hybridoma cell strain is CGMCC No.17283. Indirect ELISA valence of monoclonal antibody ascites secreted by the cell strain reaches 10<-7>, the type and the subclass of the antibody are IgG1 and kappa light chains, and the monoclonal antibody has a specific reaction with coat protein subunit of PPV. TAS-ELISA and dot-ELISA detection methods for detecting the PPV in plum trees are established on the basis of the monoclonal antibody 4A8, wherein sensitivity of the TAS-ELISA and dot-ELISA methods for detecting sick leaves reaches 20480-fold dilution and 5120-fold dilution (w/v,g/mL). The material and technical support is provided for diagnosis and detection of PPV disease in an orchard, import and export port inspection and quarantine as well as scientific prevention and control through establishment of the preparation and detection methods of the PPV monoclonal antibody.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting anti-plum pox virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Plum pox virus disease was first discovered in Bulgaria in 1915, and then spread to the entire Europe and the United States, causing serious damage to stone fruit trees in the affected areas. The pathogen Plum pox virus (Plum pox virus, PPV) has been distributed in Europe, Africa, North America, South America and Asia since Atanasoff discovered it on plum trees. Since 2001, my country has successively discovered plum pox virus in Hunan, Nanjing, and Xi'an. The Ministry of Agriculture and Rural Affairs of my country has attached great importance to it and included plum pox virus in the "List of Imported Plant Quarantine Pests of the People's Republic of China". [0003] PPV is a virus of the genus Potyvirus belonging to the family Pot...

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Application Information

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IPC IPC(8): C12N5/20C07K16/10G01N33/569G01N33/577C12R1/91
Inventor 吴建祥洪健黄悦吴佳瑜周雪平
Owner ZHEJIANG UNIV
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