虾源副溶血性弧菌PirB蛋白单克隆抗体的制备及鉴定

Preparation and Identification of Monoclonal Antibodies Against PirB Protein of Vibrio parahaemolyticus from Shrimp

用杆状病毒表达系统表达虾源副溶血性弧菌编码蛋白PirB的重组蛋白,用SDS-PAGE、Western blot对其进行鉴定,纯化后的PirB重组蛋白免疫Balb/c小鼠后进行细胞融合,通过间接ELISA方法筛选阳性杂交瘤细胞,用ELISA方法对单克隆抗体进行测定。结果发现,共筛选到7株能够稳定分泌抗PirB蛋白特异性单克隆抗体的杂交瘤细胞,小鼠腹水的效价最高效价达到1∶2.56×10^(6),制备的单克隆抗体可以特异性识别PirB蛋白,表明成功制备了虾源副溶血性弧菌PirB蛋白单克隆抗体,为虾源副溶血性弧菌引发的疾病诊断奠定了基础。

In order to develop an immunological diagnostic method for Vibrio parahaemolyticus from shrimp,the recombinant protein PirB was expressed by baculovirus expression system and identified by SDS-PAGE and Western blot.The purified PirB recombinant protein was used to immunize Balb/c mice for cell fusion.The positive hybridoma cells were screened by indirect ELISA,and the monoclonal antibodies were determined by ELISA.The results showed that seven hybridoma cells secreting monoclonal antibodies against PirB protein were screened.The highest titer of ascites was 1∶2.56×106.The monoclonal antibody can specifically recognize PirB protein,which indicates that the monoclonal antibody against PirB protein of Vibrio parahaemolyticus from shrimp has been successfully prepared,which lays a certain foundation for the detection and diagnosis of diseases caused by Vibrio parahaemolyticus from shrimp.