Hybridoma cell line stably secreting anti-pedv monoclonal antibody and its secreted antibody and application

A hybridoma cell line, monoclonal antibody technology, applied in the direction of antibodies, analytical materials, antiviral agents, etc., can solve problems such as neutralization of virus differences

Active Publication Date: 2020-05-26
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

The coronavirus S protein can induce the production of neutralizing antibodies, and the S gene mutation will inevitably lead to differences in the neutralizing antibodies induced by the S protein in terms of neutralizing the virus

Method used

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  • Hybridoma cell line stably secreting anti-pedv monoclonal antibody and its secreted antibody and application
  • Hybridoma cell line stably secreting anti-pedv monoclonal antibody and its secreted antibody and application
  • Hybridoma cell line stably secreting anti-pedv monoclonal antibody and its secreted antibody and application

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Embodiment 1

[0019] Preparation and identification of embodiment 1 anti-PEDV monoclonal antibody

[0020] 1 Materials and methods

[0021] 1.1 Cells, experimental animals, antibodies and reagents

[0022] SP2 / 0 and Vero-E6 cell lines were preserved by the Porcine Gastrointestinal Infectious Disease Innovation Team of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences; RPMI1640 culture medium (Gibco); HAT and HT medium, PEG1450, Freund's complete adjuvant, Freund's inert Complete adjuvants were purchased from Sigma; BALB / c mice were purchased from the Experimental Animal Center of our Institute.

[0023] 1.2 Culture and purification of PEDV

[0024] 1.2.1 Culture of PEDV-LNCT2 strain

[0025] After the Vero-E6 cells were overgrown to form a monolayer, the nutrient solution was discarded, washed three times with PBS (phosphate buffered saline, pH 7.2), and the PEDV-LNCT2 strain was inoculated. The dose of virus inoculation was inoculated according to the conce...

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Abstract

The invention discloses a hybridoma cell strain for stable secretion of anti-PEDV monoclonal antibody and the secreted antibody and application thereof. The hybridoma cell strain capable of stably secreting the anti-PEDV monoclonal antibody is named as 1B9, and the microbial preservation number is CGMCC No.12695. Researches prove that recognition epitope of the monoclonal antibody secreted by the hybridoma cell strain is conformational epitope. It shows through virus neutralization tests that the monoclonal antibody can effectively neutralize the strain of PEDV G2 subtype but has no neutralization activity to a strain of PEDV G1 subtype. In view of biological characteristics of the monoclonal antibody, the monoclonal antibody can be used for identification of the strains of PEDV G1 and G2 subtypes, and on this basis, a series of diagnostic methods are established. In addition, the monoclonal antibody also can be used for treating diseases caused by porcine epidemic diarrhea virus. The invention provides a novel technological means for diagnosis and treatment of PEDV G1 subtype.

Description

technical field [0001] The invention relates to a hybridoma cell strain and its secreted antibody and application, in particular to a hybridoma cell strain stably secreting anti-PEDV monoclonal antibody and its secreted antibody and application. The invention belongs to the field of biotechnology. Background technique [0002] Porcine Epidemic Diarrhea Virus (PEDV) is one of the important pathogens that cause diarrhea, emaciation, and dehydration in piglets in the pig industry, and has brought significant economic losses to the pig industry. Studies have shown that PEDV has two gene subtypes, namely G1 and G2 subtypes. Before 2010, the prevailing strains of PEDV were dominated by G1 subtype, and after 2010, the prevailing strains of PEDV were dominated by G2 subtype. Gene sequence analysis showed that the main difference between PEDV G1 and G2 subtypes was in the S gene. The S gene of the G1 subtype strain is composed of 4152nt, while the S gene of G2 is composed of 4161nt...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C12N5/20A61K39/42A61P31/14G01N33/577G01N33/569
CPCA61K2039/505C07K16/10G01N33/56983G01N33/577G01N2333/165
Inventor 冯力刘建波时洪艳陈建飞张鑫石达
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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