猪瘟病毒石门株E2蛋白的单抗制备及抗原表位鉴定  

Preparation of a monoclonal antibody against E2 glycoprotein of classical swine fever virus Shimen strain and identification of its recognition epitope

作　　者：谢文绮[1] 李得江[1] 陈宁[1] 廖迅[1] 童超[1] 方维焕[1] 

机构地区：[1]浙江大学动物预防医学研究所浙江省动物预防医学重点实验室,浙江杭州310058

出　　处：《中国兽医科学》2012年第7期661-664,共4页Chinese Veterinary Science

基　　金：浙江省科技计划项目(2011C22030);国家自然科学基金资助项目(31101793)

摘　　要：用猪瘟病毒石门株E2(SM-E2)的BC区原核表达蛋白为抗原免疫BALB/c小鼠,应用杂交瘤技术制备单抗,测定其效价并鉴定抗原表位。结果表明,制备了1株持续、稳定分泌抗SM-E2的单克隆抗体2B10,效价在1∶600～1∶1 000之间。单抗2B10能与常规SDS-PAGE转膜的SM-E2蛋白发生反应,表明其识别的是线性表位,并且该单抗只能识别石门毒株,而不能与C株疫苗及2型流行毒株发生反应。点突变E2蛋白ELISA分析结果表明,该单抗不与707、709、711、713和714位突变蛋白发生反应,表明单抗2B10识别的线性表位为707I-X-P-X-G-X-G-G714。该单抗对猪瘟病毒抗原多样性分析和致病机制研究有一定的应用价值。One hybridoma cell line secreting monoclonal antibody（McAb） 2B10 was produced by fusing mouse myeloma cells（SP2/0） with spleen cells from BALB/c mice immunized with the purified SM-E2-BC protein of classical swine fever virus（CSFV） Shimen strain. The antibody titers were 1 ： 600--1 ： 1 000. Western-blot and ELISA analysis showed that the McAb 2B10 could react with denatured SM-E2 protein, indicating that the McAb 2B10 could recognize a linear epitope. It did not react with E2 of the C strain or Group 2 strains. The McAb 2B10 could not react with mutant proteins with single alanine substitution at amino acid 707,709,711,713 and 714, further confirming that its specific recognition of a specific epitope 707^I-X-P-X-G-X-G-G^714 on E2 glycoprotein of Shimen strain. This monoclonal antibody may be helpful for the analysis of the antigenic diversity and pathogenesis studies of CSFV.

关 键 词：猪瘟病毒 囊膜糖蛋白E2 单克隆抗体 识别位点 

分 类 号：S852.659.6[农业科学—基础兽医学]