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Hybridoma cell strain, avian-like H1N1 swine influenza virus HA protein MAb, antigen epitope and application thereof

A hybridoma cell line, swine influenza virus technology, applied in the direction of antiviral immunoglobulin, immunoglobulin, biochemical equipment and methods, etc., to achieve the effect of good specificity and high titer

Active Publication Date: 2021-02-12
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is a lack of a hybridoma cell line capable of secreting HA protein MAb

Method used

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  • Hybridoma cell strain, avian-like H1N1 swine influenza virus HA protein MAb, antigen epitope and application thereof
  • Hybridoma cell strain, avian-like H1N1 swine influenza virus HA protein MAb, antigen epitope and application thereof
  • Hybridoma cell strain, avian-like H1N1 swine influenza virus HA protein MAb, antigen epitope and application thereof

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Embodiment 1

[0037] 1. Construction and identification of recombinant expression plasmid pCAGGS-HA

[0038] Virus A / swine / Zhejiang / 245 / 2013(H1N1) was isolated, identified and preserved by Harbin Veterinary Research Institute, see [Xu Huiyang, Xu Bangfeng, Chen Yan, Sui Jinyu, Yang Huanliang, Yin Hang, Yang Dawei, Qiao Chuanling, Chen Hualan. Evolutionary analysis and molecular characteristics of a H1N1 swine influenza virus. Chinese Agricultural Sciences 2015, 48(15): 3071-3078 Scientia Agriculture Sinica doi: 10.3864 / j.issn.0578-1752.2015.15.018]) inoculated 10-day-old specific pathogen-free ( SPF) chicken embryos (purchased from the Experimental Animal Center of Harbin Veterinary Research Institute), harvest the allantoic fluid after 72 hours of proliferation at 36°C, add Trizol reagent (Gibco BRL) to extract RNA, and use the universal primer uni12 (sequence: 5'-agcaaaagcagg-3 ', as shown in SEQID No.3) reverse transcription to obtain viral HA gene cDNA. Using cDNA as a template to ampl...

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Abstract

The invention provides a hybridoma cell strain, and belongs to the technical field of monoclonal antibody (MAb) preparation. The hybridoma cell strain disclosed by the invention can secrete avian-likeH1N1 swine influenza virus HA protein MAb, and the obtained MAb has the characteristics of good specificity, high titer and the like. By utilizing the MAb and combining a peptide scanning technology,the linear epitope 216VSVGSS221 located on the HA protein is obtained through screening, and the epitope is highly conservative in the avian-like H1N1 virus. The MAb and the epitope can be obtained to be used for establishing immunological diagnosis methods of the avian-like H1N1 SIV or the antibody thereof, such as ELISA, colloidal gold and the like. The HA protein epitope information of the H1N1 SIV can be perfected by obtaining the epitope information, the structure and the function of the HA protein antigen can be better understood, and theoretical and technical supports are provided forpredicting the dynamic state of virus antigenic variation and better preventing and controlling epidemic diseases.

Description

technical field [0001] The invention relates to the technical field of monoclonal antibody preparation, in particular to a hybridoma cell line, avian-like H1N1 swine influenza virus HA protein MAb, antigenic epitopes and applications. Background technique [0002] Swine influenza (SI) refers to an infectious respiratory disease of pigs caused by type A influenza virus, which has a high incidence clinically. According to the research results of SI epidemiology, it is found that H1N1, H1N2, H3N2 and other subtypes of viruses are mainly prevalent in pig herds in my country. According to the different gene sources of H1N1 virus, it is divided into classic H1N1, avian-like H1N1, human-like H1N1 and 2009 / H1N1. The avian-like type H1N1 was introduced into pigs for the first time in 1979. Now it is widespread in pigs, and sporadic infection to humans has occurred. This type of virus has important veterinary and public health significance. [0003] The genome of swine influenza vir...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/10G01N33/569
CPCC07K16/1018G01N33/56983G01N2333/11G01N2469/10
Inventor 乔传玲陈艳杨焕良陈化兰
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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