抗热毒宁注射液栀子蛋白单克隆抗体的研制与鉴定

Development and identification of monoclonal antibodies of cape jasmine proteins in Reduning injection

利用栀子脂溶性蛋白作为免疫原免疫BALB/c小鼠,取其脾细胞与SP2/0细胞进行融合,通过间接ELISA方法筛选获得1株抗栀子蛋白的单克隆抗体2H8。该单克隆抗体亚类鉴定为IgG2b,轻链为Kappa链。2H8腹水效价为1∶204 800。Western blot试验证实,2H8能与栀子蛋白反应,识别相对分子质量为58.5 kDa左右的特异性蛋白质条带。以2H8腹水作为一抗建立了斑点酶联免疫吸附试验方法(Dot-ELISA),最小检出量为19.5 ng。本研究为快速检测热毒宁注射液蛋白的试剂盒的研制奠定了基础。

Liposoluble cape jasmine proteins were used to immunize BALB/C mice. Indirect ELISA was utilized to develop one monoclonal antibody by integrating SP2/0 cells and spleen ceils from immunized BALB/C mice. The subclass of the monoclonal antibody was identified as IgG2b, with Kappa chain as its light chain. The ascite titer of 2H8 monoclonal antibody was 1 ： 204 080. Western-blot analysis proved that 2H8 reacted with cape jasmine proteins to identify specific liposoluble protein with molecuar weight of around 58. 5 kDa. Dot-ELISA was established with 2H8 ascites as the primary antibody, showing the minimum detectable amount of 19. 5 ng. This study lays a foundation for the development of protein kits of Reduning injection.