期刊论文

Production and detection of monoclonal antibodies against recombinant nucleoprotein of TGEV

中文

中国兽医科学

1673-4696

2018

48

1

13-18

10.16656/j.issn.1673-4696.2018.0003

2016C32072:公益性行业(农业)科研专项 20162012A07:杭州市农业科研主动设计项目

本校为第一机构

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IgG1,腹水的间接ELISA效价均为1 ∶ 10~8。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 80...