流行性乙型脑炎病毒E蛋白单克隆抗体的制备与鉴定

Development and identification of the monoclonal antibodies against recombinant JEV E protein

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 of public health importance. The envelope glycoprotein of JEV is associated with viral attachment and fusion with host cells, virus hemagglutination ability, cellular tropism, viral virulence, and induction of protective immune response. Monoclonal anti- body （McAb） 5D4 against Japanese encephalitis virus E protein of 8A14-14-2 strain were developed by fu- sing myeloma cell line SP2/0 with splenocytes of BALB/c mice immunized with prokaryotic expressed E protein. The monoclonal antibody was determined to be IgG2a subtype, and its light chain was n chain. Western blot analysis indicated that the monocl.onal antibody could specifically react with JEV E protein, and it may be used as an valuable tools for diagnosis of JEV.