抗传染性支气管炎病毒S1蛋白单克隆抗体的制备与鉴定  

Preparation and Identification of Monoclonal Antibody Against IBV S1 Protein

作　　者：李佳楠 杜林 周景明 祁艳华 马强 张改平 王爱萍 LI Jia-nan;DU Lin;ZHOU Jing-ming;Qi Yan-hua;MA Qiang;ZHANG Gai-ping;WANG Ai-ping(College of Life Sciences,Zhengzhou University,Zhengzhou,Henan,450000,China;College of Life Sciences,Northeast Forestry University,Harbin,Heilongjiang,150040,China)

机构地区：[1]郑州大学生命科学学院,河南郑州450001 [2]东北林业大学生命科学学院,黑龙江哈尔滨150040

出　　处：《动物医学进展》2021年第3期17-20,共4页Progress In Veterinary Medicine

基　　金：“十三五”国家重点研发计划项目(2016YFD0500800)。

摘　　要：选取传染性支气管炎病毒(Infectious bronchitis virus, IBV)M41毒株S1蛋白的重要抗原区,RT-PCR扩增选定的S1基因并构建pET-28a-S1重组表达载体,经IPTG诱导表达后,SDS-PAGE以及Western blot结果显示重组S1蛋白表达正确。以纯化的S1蛋白作为免疫原免疫Balb/c小鼠,采用杂交瘤技术制备并获得了抗IBV M41 S1蛋白的单克隆抗体3D9。免疫过氧化酶单层试验(immunoperoxidase monolayer assay, IPMA)结果显示,单克隆抗体3D9可与IBV M41毒株反应。成功表达了截短的IBV S1蛋白并获得了能识别IBV M41毒株的单克隆抗体,为IBV检测方法的建立奠定了基础。In this study,we selected the important antigen region of IBV M41 S1 protein,the S1 gene was amplified by RT-PCR and the pET-28a-S1 recombinant expression vector was constructed.After induction by IPTG,the correct expression of recombinant S1 protein was verified by SDS-PAGE and Western blot.The purified S1 protein was used as an immunogen to immunize Balb/c mice.A monoclonal antibody 3D9 against IBV M41 S1 protein was prepared by cell hybridization.IPMA results showed that the monoclonal antibody 3D9 could react with the IBV M41 strain.This test successfully expressed the IBV S1 protein and prepared a monoclonal antibody that can recognize the IBV M41 strain,which laid a foundation for the detection of IBV.

关 键 词：传染性支气管炎病毒 S1蛋白 单克隆抗体 免疫过氧化酶单层试验 

分 类 号：S852.659.4[农业科学—基础兽医学]