Hybridoma cell strain for secreting duck type-1 hepatitis A virus subtype monoclonal antibody

A technology of hybridoma cell lines and monoclonal antibodies, applied in antiviral immunoglobulin, analytical materials, biochemical equipment and methods, etc.

Active Publication Date: 2016-03-02
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
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Problems solved by technology

[0009] The technical problem to be solved in the present invention is to solve the subtype monoclonal antibody that does not have duck type 1 hepatitis A virus at present, can not be used for the subtype monoclonal antibody of duck type 1 hepatitis A virus In view of the problems of type detection and trea...

Method used

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Embodiment Construction

[0049] Preferred embodiments of the present invention are described below, and it should be understood that the preferred embodiments described here are only used to illustrate and explain the present invention, and are not intended to limit the present invention.

[0050] The preparation method of the subtype monoclonal of duck type 1 hepatitis A virus is realized in the following steps:

[0051] 1. Preparation of immunogen

[0052] That is, the duck type 1 hepatitis A virus subtype is inactivated and purified by ultracentrifugation, and used as an immune antigen. Use duck fibroblasts to propagate 500mL duck type 1 hepatitis A virus subtype, collect the cell culture medium, centrifuge to remove the cell sediment, inactivate with 0.3% formaldehyde, then add an appropriate amount of chloroform for overnight treatment, and centrifuge at 10000rpm for 15min , collect the supernatant, then centrifuge at 25,000rpm for 1h to remove the precipitate, and then centrifuge at 40,000rpm...

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Abstract

The invention discloses a hybridoma cell 1A2 for secreting a duck type-1 hepatitis A virus subtype monoclonal antibody. The preservation number is CGMCC NO.:10420. After a mouse is immunized three times by purified duck type-1 hepatitis A virus subtype inactivated totivirus antigens, VP1 protein is used for boosting immunization; the mouse spleen taking cells with the highest duck type-1 hepatitis A virus subtype antibody titer are integrated with SP2/0 bone marrow cells, and the hybridoma cell 1A2 for resisting the duck type-1 hepatitis A virus subtype VP1 protein is screened out. The monoclonal antibody 1A2 obtained through the method has the high specificity, high titer, good stability and the like and can be applied to preparing a duck type-1 hepatitis A virus subtype antibody detection agent.

Description

technical field [0001] The invention relates to the field of etiology of duck type 1 hepatitis A virus subtype, in particular to a hybridoma cell strain secreting duck type 1 hepatitis A virus subtype monoclonal antibody and application thereof. Background technique [0002] Duck hepatitis A virus type 1 (duckhepatitisAvirus1, DHAV-1) is a picornavirus family ( Pacornaviridae ) Avian Hepavirus ( Avihepatovirus One of the members of ), it mainly affects ducklings about 6 weeks old. The clinical symptoms are acute onset, short course of disease, high lethality and opisthotonos in dead ducks, and the gross lesion features are hepatomegaly and hemorrhage. [0003] In 2005, Guérin et al. isolated duck hepatitis A virus type 1 from muscovy ducks showing pancreatitis and encephalitis, and the liver of the affected ducks did not appear swollen and hemorrhagic lesions. [0004] In September 2011, in Fujian, Zhejiang, Shanghai, Guangdong and other provinces of my country, 10-30-day...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/10G01N33/577G01N33/569
Inventor 傅秋玲傅光华黄瑜程龙飞施少华万春和陈红梅林建生陈珍陈翠腾朱春华
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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