猪瘟病毒E2蛋白A/D区单抗的制备及其抗原表位的初步研究  

Preparation and Identification of Monoclonal Antibodies against the Truncated E2 Protein of Classical Swine Fever Virus

作　　者：张春玲[1] 宗先伟 杨文超[1] 张敬梅[1] 彭明义[1] 程宝艳[3] 余为一[1] 

机构地区：[1]安徽农业大学动物科技学院,合肥230036 [2]山东六和集团有限公司,德州253000 [3]安徽省农业科学院畜牧兽医研究所,合肥230031

出　　处：《中国畜牧兽医》2009年第9期50-54,共5页China Animal Husbandry & Veterinary Medicine

基　　金：安徽省"十五"科技攻关项目(01013003)资助

摘　　要：应用分子克隆技术将猪瘟病毒E2蛋白部分基因插入到载体pGEX-4T-1中构建重组质粒pGEX-4T-E(A),在大肠杆菌Rosetta中表达融合蛋白GST-E(A),以纯化的融合蛋白免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞SP2/0进行融合,经克隆化和间接ELISA筛选,获得了C3、A1两株稳定分泌抗猪瘟病毒E2蛋白单克隆抗体的杂交瘤细胞株,ELISA结果显示,其腹水效价为1∶128000和1∶256000。Western blotting结果证实,单克隆抗体C3和A1能与猪瘟病毒(classical swinefever virus,CSFV)发生特异性的反应,表明该单抗是针对猪瘟病毒E2蛋白的保守线性表位。A truncated gene encoding the major antigenic domains of E2 protein of classical swine fever virus （CSFV） was amplified and cloned into pGEX-4T-1 expression vector to obtain recombinant pGEX-4T-E（A）. The recombinant GST-E（A） protein expressed in E. coli Rosetta. BALB/c mice was immunized with purified GST-E（A）protein as antigen and mouse splenic cells were fused with SP2/0 cells. Hybridoma cells were screened by ELISA and subcloned. Two hybridoma cells C3 and A1 secreting anti-CSFV monoclonal antibodies were obtained. The EI.ISA showed the antibody titres were 1 ： 128000 and 1 ： 256000. Western blotting indicated that C3 and A1 reacted with CSFV specially. These results demonstrated that the McAbs were specific reagent for CSFV, and recognized a linear conserved epitope on the E2 protein.

关 键 词：猪瘟病毒 E2蛋白 单克隆抗体 鉴定 

分 类 号：S859.797[农业科学—临床兽医学]