鸡传染性支气管炎病毒（poly100）S1蛋白单克隆抗体的制备与鉴定

Production and characterization of monoclonal antibodies to (poly100)S1 protein of avian infectious bronchitis virus.

作者信息

Hu J Q, Li Y F, Guo J Q, Shen H G, Zhang D Y, Zhou J Y

机构信息

Laboratory of Virology and Immunology, Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou 310029, China.

出版信息

Zoonoses Public Health. 2007;54(2):69-77. doi: 10.1111/j.1863-2378.2007.01030.x.

DOI:10.1111/j.1863-2378.2007.01030.x

PMID:17348910

Abstract

Fragments within S1 genes ((poly100)S1) of infectious bronchitis virus (IBV) strains ZJ971, M41 and SC021202 (SC) were subcloned into a prokaryotic expression vector and expressed in Escherichia coli. Monoclonal antibodies (mAbs) against the recombinant (poly100)S1 proteins were produced, characterized and used to analyse epitopes on the S1 subunit of IBV. Nine mAbs raising from the three (poly100)S1 proteins recognized five different epitopes of the S1 subunit, designated as S1-A, B, C, D and E. Epitopes S1-C and S1-D are common for the three IBV strains, while S1-A and S1-B exist on ZJ971 and M41 strains, and S1-E was a strain-specific epitope for SC strain. Immunocytochemistry indicated that all the mAbs to the (poly100)S1 proteins can react with the homologous S1 glycoprotein expressed in Vero cells. Moreover neutralization test demonstrated that only mAbs 6E2, 4F9 and 6G4 had neutralization activity for the homologous IBV. These mAbs to (poly100)S1 protein were potential candidates for detecting and distinguishing IBV strains, and also used to examine antigenic variation of the S1 protein.

摘要

将传染性支气管炎病毒（IBV）毒株ZJ971、M41和SC021202（SC）的S1基因片段（（poly100）S1）亚克隆到原核表达载体中，并在大肠杆菌中表达。制备、鉴定了针对重组（poly100）S1蛋白的单克隆抗体（mAb），并用于分析IBV S1亚基上的表位。从三种（poly100）S1蛋白中产生的9种mAb识别出S1亚基的5种不同表位，分别命名为S1 - A、B、C、D和E。表位S1 - C和S1 - D是三种IBV毒株共有的，而S1 - A和S1 - B存在于ZJ971和M41毒株上，S1 - E是SC毒株特有的表位。免疫细胞化学表明，所有针对（poly100）S1蛋白的mAb都能与Vero细胞中表达的同源S1糖蛋白发生反应。此外，中和试验表明，只有mAb 6E2、4F9和6G4对同源IBV具有中和活性。这些针对（poly100）S1蛋白的mAb是检测和区分IBV毒株的潜在候选物，也用于检测S1蛋白的抗原变异。