抗蓝舌病病毒VP6和VP7蛋白单克隆抗体的制备及鉴定  

Preparation and identification of monoclonal antibodies against VP6 and VP7 of bluetongue virus

作　　者：魏鹏[1] 孙恩成[1] 刘霓红[1] 杨涛[1,2] 徐青元[1] 秦永丽[1] 赵晶[1] 冯瑜菲[1,2] 王文世[1] 张翠云[1,3] 吴东来[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所、兽医生物技术国家重点实验室/农业部兽医公共卫生重点开放实验室,黑龙江哈尔滨150001 [2]东北农业大学动物医学学院,黑龙江哈尔滨150030 [3]湖南农业大学动物医学学院,湖南长沙410128

出　　处：《中国预防兽医学报》2012年第5期412-414,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：公益性行业(农业)科研专项经费(20120217);国家863项目(2011AA10A212)

摘　　要：为制备针对蓝舌病病毒(BTV)的单克隆抗体(MAb),本研究利用血清型1型BTV(BTV1)免疫BALB/c鼠,将其脾淋巴细胞与SP2/0进行融合,并用BTV1包被ELISA板,通过间接ELISA方法筛选出3株稳定分泌抗BTV1的MAb的杂交瘤细胞株(2B10、3D4和4H8)。利用表达BTV1主要蛋白的真核表达重组质粒转染BHK-21后,对所制备的杂交瘤细胞株上清进行间接免疫荧光(IFA)以及western blot鉴定,结果显示:2B10和4H8与VP7蛋白反应,而3D4与VP6蛋白反应。同时,IFA鉴定结果进一步表明,3株MAb与24个血清型的BTV均可以发生反应。本研究制备的MAb为建立BTV免疫学检测方法和相关病毒蛋白的功能研究奠定了基础。To prepare a monoclonal antibody （MAb） against Bluetongue virus （BTV）, BALB/c mice were immunized with BTV1 and the immunized mice spleen myeloma cells were fused with SP2/0, then three MAbs （2B10, 3D4 and 4H8） against BTV were screened by indirect ELISA coated with BTV1. The MAbs were identified by indirect immunofluorescence assay （IFA） and western blot, the results showed that the MAbs of 2B10 and 4H8 were reacted with BTV VP6 and 3D4 with VP7 protein, which were expressed in transfected BHK-21 cells with eukaryocyte recombinant plasmids. Furthermore, IFA identification indicated that all the MAbs were reacted to BTV 1 to 24 serotypes. The MAbs prepared in this study are useful tools for developing immunological diagnostic techniques of BTV and investigating proteins related to BTV.

关 键 词：蓝舌病病毒 单克隆抗体 VP6蛋白 VP7蛋白 

分 类 号：S852.65[农业科学—基础兽医学]