Hybridoma strain capable of secreting monoclonal antibodies for preventing citrus tristeza virus and monoclonal antibody applications thereof

A technology of citrus decay virus and hybridoma cell line, which is applied in antiviral immunoglobulin, analytical materials, biochemical equipment and methods, etc., and can solve problems such as low efficiency

Active Publication Date: 2015-03-11
ZHEJIANG UNIV +1
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  • Claims
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Problems solved by technology

At present, only low-efficiency electron microscope observation, RT-PCR method, nucleic acid hybridization, RT-PCR and other methods are used to detect viruses in small samples.

Method used

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  • Hybridoma strain capable of secreting monoclonal antibodies for preventing citrus tristeza virus and monoclonal antibody applications thereof
  • Hybridoma strain capable of secreting monoclonal antibodies for preventing citrus tristeza virus and monoclonal antibody applications thereof

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Embodiment Construction

[0011] The hybridoma cell line secreting anti-citrus decay virus monoclonal antibody was deposited on July 3, 2014 in the General Microbiology Center of the Chinese Microbiological Culture Collection Management Committee, Institute of Microbiology, Chinese Academy of Sciences, address: 1 Beichen West Road, Chaoyang District, Beijing No. 3 Courtyard, Zip Code: 100101, and the preservation number is CGMCC No.9335, which can secrete monoclonal antibodies against citrus decay virus.

[0012] The indirect ELISA titer of the ascitic fluid monoclonal antibody against citrus recession virus secreted by the hybridoma cell line reaches 10 -8 , the antibody type and subclass are IgG1, kappa chain, the monoclonal antibody can have a specific immune binding reaction with the coat protein subunit of citrus decay virus, and the sensitivity of the monoclonal antibody to detect diseased leaves reaches 1:10240 times dilution.

[0013] The monoclonal antibody against citrus decay virus secreted ...

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Abstract

The invention discloses a hybridoma strain capable of secreting monoclonal antibodies for preventing citrus tristeza virus and monoclonal antibody applications thereof. The coat protein gene of citrus tristeza virus (CTV) is cloned, the CTV coat protein is expressed by utilizing a prokaryotic expression system, then the purified coat protein is taken as an antigen to immunize a BALB / c mouse, through the steps of cell fusion, screening, and cloning, a hybridoma strain 14H11, which can stably pass the characteristics to the next generation and secret monoclonal antibodies for preventing CTV, is obtained, and the preservation number of the hybridoma strain is CGMCC No.9335. The ascites indirect ELISA titer of the 14H11 monoclonal antibody can reach 10-8, and the antibody type and subtype are IgG1, kappa chain. The established dot-ELISA method can precisely, specifically, and sensitively detect the CTV from the orange species samples. The preparation of CTV monoclonal antibody and establishment of a detection method utilizing the monoclonal antibody provide technological and material support for the diagnosis, detection, and scientific prevention and control on CTV.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a hybridoma cell strain secreting anti-citrus recession virus monoclonal antibody and the application of the monoclonal antibody. Background technique [0002] Citrus tristeza virus (CTV) is widely distributed in citrus producing areas all over the world, and the decay disease caused by it is one of the most harmful citrus diseases. The virus is mainly transmitted by aphids and infected scions. CTV has obvious strain differentiation. Except for a few strains that do not produce visible symptoms on some citrus varieties, most strains can produce obvious symptoms on citrus. The common symptoms can be divided into three types, namely the fast-declining type , stem pit type and seedling yellow type. The fast-declining symptoms are mainly manifested in susceptible citrus such as sweet oranges that use sour oranges as rootstocks; the occurrence of stem pitting symptoms has nothing to do w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/10G01N33/577G01N33/569C12R1/91
Inventor 吴建祥洪健陈浙周雪平刘震刘金香周常勇
Owner ZHEJIANG UNIV
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