猪传染性胃肠炎病毒N蛋白单克隆抗体的制备及鉴定  

Production and detection of monoclonal antibodies against recombinant nucleoprotein of TGEV

作　　者：王赛[1,2] 苏菲 袁秀芳[2] 徐丽华[2] 李军星[2] 余兴龙[1] 

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128 [2]浙江省农业科学院畜牧兽医研究所,浙江杭州310021

出　　处：《中国兽医科学》2018年第1期13-18,共6页Chinese Veterinary Science

基　　金：公益技术研究农业项目(2016C32072);杭州市农业科研主动设计项目(20162012A07)

摘　　要：以纯化的猪传染性胃肠炎病毒（TGEV）重组N蛋白免疫BALB/c小鼠,按照常规方法融合、筛选,最终获得2株抗TGEV N蛋白的杂交瘤细胞系,分别命名为2G7和2C2。2株单抗亚类均为Ig G1,腹水的间接ELISA效价均为1∶108。2株杂交瘤细胞连续培养20代,各代次的上清效价保持不变,均为1∶12 800,说明2株细胞分泌抗体的能力稳定。通过间接免疫荧光发现,制备的2株单克隆抗体能与感染TGEV的细胞发生特异性反应,在胞浆和细胞膜上有亮绿色荧光。结果表明,所制备的2株TGEV N蛋白的特异性单克隆抗体细胞株遗传稳定,分泌的抗体敏感性高,为TGEV病原检测奠定了基础。BALB/c mice were immunized with purified recombinant N protein（rNp） of porcine trans- missible gastroenteritis virus（TGEV）.Cell fusion and screening were carried out according to standard methods. Finally,two hybridoma cell lines against TGEV NP were obtained and named as 2G7 and 2C2,re- spectively. The isotypes of the two monoclonal antibodies were found to be IgGi. Ascites titer of two monoclonal antibodies was i： 108. The result of indirect ELISA showed that the antibody titers of super- natant of the 20th passage of cells were 1：12 800,indicating stable and strong antibody secreting ca- pacity of the two hybridoma. Moreover,the result of immunofluorescent analysis showed high specificity of the two monoclonal antibodies against TGEV. These results demonstrated that the two monoclonal anti- bodies prepared in our study had high sensitivity,and might provide valuable tools for accurate and rapid diagnosis of TGEV.

关 键 词：传染性胃肠炎病毒 重组N蛋白 单克隆抗体 

分 类 号：S852.659.2[农业科学—基础兽医学]