论文部分内容阅读
流行性乙型脑炎病毒(JEV)是一种严重危害人畜健康的虫媒病毒。表面囊膜蛋白(E蛋白)是该病毒的主要结构蛋白。本研究利用原核表达的乙型脑炎病毒SA14-14-2株结构蛋白E蛋白作为免疫原,免疫6周龄BALB/c小鼠,采用淋巴细胞杂交瘤技术进行融合,经有限稀释法获得1株特异性针对JEV E蛋白的杂交瘤细胞,命名为5D4,经测定5D4单抗亚类属于IgG2a,轻链为κ链。经Western blot证实所得杂交瘤细胞分泌的抗体可特异地与JEV E蛋白结合。结果表明,所制备的抗JEV E蛋白的单抗对病原检测具有很高的特异性,为JEV准确快速的病原诊断和JEV感染相关的基础性研究提供了物质基础。
Japanese encephalitis virus (JEV) is an arbovirus that seriously endangers the health of humans and animals. Surface envelope protein (E protein) is the virus’s major structural protein. In this study, BALB / c mice of 6 weeks old were immunized with the structural protein E of prokaryotic expression of Japanese encephalitis virus SA14-14-2 strain as the immunogen and fused by lymphocyte hybridoma technique. Strains of hybridomas specific for the JEV E protein, designated 5D4, were determined to belong to the IgG2a subclass of the 5D4 monoclonal antibody and the kappa chain of the light chain. Antibodies secreted by the resulting hybridoma cells were confirmed by Western blot to specifically bind to the JEV E protein. The results showed that the McAb against JEV E protein was highly specific for pathogen detection and provided the material basis for the accurate and rapid pathogen diagnosis of JEV and the basic research related to JEV infection.