Hybridoma cell strain capable of secreting tomato mottle mosaic virus monoclonal antibodies, and application of monoclonal antibodies

A hybridoma cell line, mottled flower technology, applied in antiviral immunoglobulin, analytical materials, biological material analysis and other directions, to achieve the effect of low technical requirements

Active Publication Date: 2020-06-12
ZHEJIANG UNIV
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However, the establishment of serological methods

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  • Hybridoma cell strain capable of secreting tomato mottle mosaic virus monoclonal antibodies, and application of monoclonal antibodies
  • Hybridoma cell strain capable of secreting tomato mottle mosaic virus monoclonal antibodies, and application of monoclonal antibodies
  • Hybridoma cell strain capable of secreting tomato mottle mosaic virus monoclonal antibodies, and application of monoclonal antibodies

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Embodiment Construction

[0016] The hybridoma cell line 2D6 secreting tomato mottle mosaic virus monoclonal antibody was deposited on November 4, 2019 in the General Microbiology Center of China Microbiology Culture Collection Management Committee, Institute of Microbiology, Chinese Academy of Sciences, and the preservation number is CGMMCNO.18846. Monoclonal antibody against mottle mosaic virus.

[0017] A kind of described hybridoma cell secretes tomato mottle mosaic virus monoclonal antibody, and this monoclonal antibody ascitic fluid indirect ELISA titer reaches 10 -8 , the antibody type and subclass are IgG 2a , κ light chain, which has a specific immune reaction with the 17.5kDa coat protein of tomato mottle mosaic virus, using the ACP-ELISA and dot-ELISA methods established by this monoclonal antibody to detect crude extracts of diseased leaf tissues infected with tomato mottle mosaic virus The sensitivity reaches 1:40,960 and 1:20,480 times dilution (w / v, g / mL) respectively.

[0018] Monoclo...

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Abstract

The invention discloses a hybridoma cell strain capable of secreting an anti-tomato mottle mosaic virus (ToMMV) monoclonal antibody, and application of the monoclonal antibody of the hybridoma cell strain. According to the invention, purified ToMMV virus particles are used as antigens to immunize BALB/c mice, and through cell fusion, screening and cloning, a hybridoma cell strain 2D6 capable of secreting an anti-ToMMV monoclonal antibody is obtained, wherein the preservation number of the hybridoma cell strain 2D6 is CGMMC NO.18846, the ascites indirect ELISA titer of the monoclonal antibody secreted by the cell strain reaches 10<-8>, and the monoclonal antibody secreted by the cell strain performs a specific immune reaction with ToMMV, but not reacts with Tobacco mosaic virus (TMV) and tomato mosaic virus (ToMV) which belong to the same genus; an ACP-ELISA detection method, a dot-ELISA detection method and a Tissue print-ELISA detection method for detecting ToMMV in a tomato plant areestablished by utilizing the 2D6 monoclonal antibody, wherein the sensitivity of the ACP-ELISA method and the dot-ELISA method for detecting the diseased leaves reaches 1:40, 960 times dilution (w/v,g/mL) and 1:20 and 480 times dilution (w/v, g/mL) respectively; and the creation of the hybridoma cell strain capable of secreting high-quality ToMMV monoclonal antibody and the establishment of thevirus serological detection method provide material and technical support for ToMMV diagnosis and detection, epidemiological investigation and scientific prevention and control.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting tomato mottle mosaic virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Tomato mottle mosaic virus (ToMMV) is a new species of Tobamovirus in the family Viraviridae, and its genome is positive-sense single-stranded RNA. The genome of ToMMV contains four open reading frames. Like tobacco mosaic virus, it expresses four proteins, encoding a 126kD and a 183kD replication-required protein, and also encoding a 30kD motor protein and a 17.5kD coat protein. In 2013, the new virus was first discovered on Mexican tomatoes, and then the virus was detected on tomatoes in the United States, Israel, Brazil, Spain and other places. Tomatoes in Yunnan and Hainan provinces in my country were also infected by the virus. [0003] ToMMV can harm a variety of Solanaceae crops, and the main symptoms of the disease ar...

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Application Information

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IPC IPC(8): C12N5/20C07K16/10G01N33/569G01N33/577
CPCC07K16/10C07K2317/33C07K2317/35G01N33/56983G01N33/577G01N2333/08
Inventor 吴建祥郭丽倩吴佳瑜钱亚娟周雪平
Owner ZHEJIANG UNIV
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