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A kind of hybridoma cell line, poultry type h1n1 swine influenza virus ha protein mab, antigenic epitope and application

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

Active Publication Date: 2021-07-20
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A kind of hybridoma cell line, poultry type h1n1 swine influenza virus ha protein mab, antigenic epitope and application
  • A kind of hybridoma cell line, poultry type h1n1 swine influenza virus ha protein mab, antigenic epitope and application
  • A kind of hybridoma cell line, poultry type h1n1 swine influenza virus ha protein mab, antigenic epitope and application

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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 of the invention can secrete the HA protein MAb of poultry-like H1N1 swine influenza virus, and the obtained MAb has the characteristics of good specificity, high potency and the like. The present invention uses the above-mentioned MAb, combined with peptide scanning technology, to screen and obtain a linear epitope located on the HA protein 216 VSVGSS 221 , this epitope is highly conserved among avian H1N1 viruses. The acquisition of MAb and antigenic epitope can be used to establish immunological diagnostic methods for avian-like H1N1 SIV or its antibody, such as ELISA, colloidal gold, etc. The acquisition of epitope information is helpful to improve the epitope information of H1N1 SIV HA protein, to better understand the structure and function of HA protein antigen, to predict the dynamics of virus antigenic variation, and to better prevent and control the epidemic. Theoretical and technical support.

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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Patent Type & Authority Patents(China)
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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