Hybridoma cell line secreting tomato mottle mosaic virus monoclonal antibody and its application

A technology of hybridoma cell lines and mottled flowers, which is applied in antiviral immunoglobulin, analytical materials, biological material analysis, etc., to achieve the effect of low technical requirements

Active Publication Date: 2021-06-04
ZHEJIANG UNIV
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Problems solved by technology

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

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  • Hybridoma cell line secreting tomato mottle mosaic virus monoclonal antibody and its application
  • Hybridoma cell line secreting tomato mottle mosaic virus monoclonal antibody and its application
  • Hybridoma cell line secreting tomato mottle mosaic virus monoclonal antibody and its application

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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 the Chinese Microbiological Culture Collection Management Committee, Institute of Microbiology, Chinese Academy of Sciences, and the preservation number is CGMMCNO.18846. It can secrete anti-tomato 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...

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Abstract

The invention discloses a hybridoma cell line secreting anti-tomato mottle mosaic virus (ToMMV) monoclonal antibody and the application of the monoclonal antibody. BALB / c mice were immunized with purified ToMMV virus particles as antigen, and a hybridoma cell line 2D6, which can secrete anti-ToMMV monoclonal antibody, was obtained through cell fusion, screening and cloning, and its preservation number was CGMMC NO.18846. The cell line secretes monoclonal antibody ascitic fluid indirect ELISA titer up to 10 ‑8 , which has a specific immune reaction with ToMMV, but not with the same genus of tobacco mosaic virus (Tobacco mosaic virus, TMV) and tomato mosaic virus (Tomato mosaic virus, ToMV). ACP‑ELISA, dot‑ELISA and Tissue print‑ELISA detection methods for detecting ToMMV in tomato plants were established using 2D6 monoclonal antibody, among which the sensitivity of ACP‑ELISA and dot‑ELISA in detecting diseased leaves reached 1:40,960 and 1:20,480 times, respectively Dilution (w / v, g / mL). The creation of ToMMV-secreting high-quality monoclonal antibody hybridoma cell lines and the establishment of serological detection methods for the virus provide material and technical support for the diagnosis and detection, epidemiological investigation, and scientific prevention and control of ToMMV.

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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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