DOI number:10.16372/j.issn.1004-6364.2021.05.005
Affiliation of Author(s):Yangzhou University
Journal:China poultry
Key Words:infectious bronchitis virus, N protein, monoclonal antibody
Abstract:The objective of this study was to prepare broad-spectrum monoclonal antibodies that could react with a variety of genotypes of infectious bronchitis virus (IBV), and the response spectrum could cover the dominant genotypes currently prevalent in China. The N gene of CK/CH/JS/2010/12 strain of QX-type was cloned by RT-PCR, and then inserted into prokaryotic expression vector pET-32a(+) and pGEX-6P-1, respectively, to construct recombinant plasmids pET-N and pGEX-N. Two recombinant fusion proteins, His N and GST-N, were prokaryotic expressed in Escherichia coli and purified by affinity chromatography method. Balb/c mice were immunized with recombinant protein GST- N. Spleen cells of immunized mice were harvested and fused with myeloma cells SP2/0 by lymphocyte hybridoma technique. The positive hybridoma cells were screened by indirect ELISA method established by using recombinant protein His-N as detection antigen. After screening and subcloning, 12 hybridoma cell lines were obtained. Western blot results showed that 8 of the monoclonal antibodies could react specifically with all the genotypes of IBV tested in this study, and the other 4 antibodies could react with all strains other than 4/91 of 793/B-type. In conclusion, broad- spectrum monoclonal antibodies against IBV were successfully obtained, which laid a foundation for further research to establish a universal method for detecting IBV antigen.
Co-author:Jia Zhao,Mengjiao Guo,Min Li,Chengcheng Zhang,Yantao Wu
First Author:Xubin Du
Indexed by:Research Atricle
Correspondence Author:Xiaorong Zhang
Document Code:1004-6364（2021）05-27-06
Discipline:Agricultural science
First-Level Discipline:Veterinary Medicine
Document Type:J
Volume:43
Issue:5
Page Number:27-32
Translation or Not:no
Date of Publication:2021-05-15