Hybridoma cell strain for stable secretion of anti-PEDV monoclonal antibody and secreted antibody and application thereof

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: 2017-02-08
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
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  • Claims
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AI Technical Summary

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 strain for stable secretion of anti-PEDV monoclonal antibody and secreted antibody and application thereof
  • Hybridoma cell strain for stable secretion of anti-PEDV monoclonal antibody and secreted antibody and application thereof
  • Hybridoma cell strain for stable secretion of anti-PEDV monoclonal antibody and secreted antibody and application thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

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

[0020] 1Materials and methods

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

[0022] SP2 / 0 and Vero-E6 cell lines are preserved by the pig digestive tract infectious diseases innovation team of Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences; RPMI1640 culture medium (Gibco); HAT and HT medium, PEG1450, Freund's complete adjuvant, Freund Complete adjuvants were purchased from Sigma; BALB / c mice were purchased by our laboratory animal center.

[0023] 1.2 Cultivation and purification of PEDV

[0024] 1.2.1 Culture of PEDV-LNCT2 strain

[0025] After Vero-E6 cells overgrown to form a monolayer, the nutrient solution was discarded, washed three times with PBS (phosphate buffered solution, pH 7.2), and inoculated with the PEDV-LNCT2 strain. The virus inoculation dose was inoculated at a concentration of 5%, and the virus was inoculated with pancreatin at th...

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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 present invention relates to a hybridoma cell strain and its secreted antibodies and applications, in particular to a hybridoma cell strain which stably secretes anti-PEDV monoclonal antibodies and its secreted antibodies and applications. The present invention belongs to the field of biotechnology. Background technique [0002] Porcine Epidemic Diarrhea Virus (PEDV) is one of the important pathogens that cause piglet diarrhea, weight loss and dehydration in the pig industry, and it brings significant economic losses to the pig industry. Studies have shown that PEDV has two gene subtypes, namely G1 and G2 subtypes. Before 2010, the predominant PEDV strain was G1 subtype, and after 2010, the predominant PEDV strain was G2 subtype. Gene sequence analysis showed that the main difference between PEDV G1 and G2 subtypes lies in the S gene. The S gene of the G1 subtype strain is composed of 4152 nt, while the S gene of G2 is composed of 4161 nt, which incre...

Claims

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Application Information

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Patent Type & Authority Applications(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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