犬瘟热病毒截短P蛋白单克隆抗体的制备与鉴定  

Preparation and Identification of Monoclonal Antibodies against Recombinant Truncated P Protein of Canine Distemper Virus

作　　者：袁维峰[1] 贾红[1] 于萍萍[1,2] 侯绍华[1] 郭晓宇[1] 史利军[1] 赵占中[1] 朱鸿飞[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]山东农业大学动物科技学院,山东泰安271017

出　　处：《中国畜牧兽医》2013年第4期22-25,共4页China Animal Husbandry & Veterinary Medicine

基　　金：中央级公益性科研院所基本科研业务费专项资金项目(2011js-3;2010js-1;2010js-2);2012年农业部动物疫情监测与防治项目

摘　　要：为制备针对犬瘟热病毒(canine distemper virus,CDV)P1蛋白的单克隆抗体(McAb),本研究利用重组犬瘟热病毒P1蛋白免疫BALB/c小鼠,并将其脾淋巴细胞与SP2/0进行融合,用CDV包被ELISA板,通过间接ELISA法筛选出两株稳定分泌抗CDV-P1的杂交瘤细胞株(E9E4C10、E9F8D9)。间接ELISA检测腹水效价均为1∶106,亚类鉴定结果均为IgG2b。Western blotting和间接免疫荧光(IFA)分析结果表明,两株单克隆抗体均能与重组P1蛋白和CDV发生反应;ELISA叠加试验增殖结果表明,两株单克隆抗体识别的抗原表位不同。本研究制备的特异性抗CDV-P1的单克隆抗体为建立CDV免疫学检测方法和P蛋白的功能研究奠定了基础。The assay was aimed to prepare monoclonal antibodies （McAbs） against P1 protein of canine distemper virus （CDV）. BALB/c mice were immunized with P1 protein and the immunized mice spleen myeloma cells were fused with SP2/0. Two hybridoma cell lines of E9E4C10 and E9F8D9 that stably produced antibodies against protein were identified by ELISA. The antibody titres of ascites for the hybridoma lines were both 1∶106. The subtypes of monoclonal antibodies were both identified as IgG2b. Western blotting and IFA showed that the McAbs specifically recognized certain antigenic epitopes of P1 protein of CDV. Additive index of ELISA competitive binding assay indicated that the two monoclonal antibodies were specific for different epitopes. The McAbs prepared in this study were useful tools for developing immunological diagnosis techniques of CDV and investing P protein function.

关 键 词：犬瘟热病毒 重组P蛋白 单克隆抗体 鉴定 

分 类 号：S852.655[农业科学—基础兽医学]