Ruan Xindi, Huang Shaomei, Shao Lin, Ye Jing, Chen Zheng, Chen Huanchun, Cao Shengbo
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University , Wuhan, Hubei, China .
Monoclon Antib Immunodiagn Immunother. 2013 Dec;32(6):382-5. doi: 10.1089/mab.2013.0041.
Japanese encephalitis (JE) is one of the most prevalent global viral encephalitis viruses. The functions of JEV (virus) NS4B protein are still under investigation. In our study, NS4B was expressed in Escherichia coli and purified by dialysis. Two clones of monoclonal antibodies (MAbs 1B1 and 1C3) against NS4B protein were generated and their characterizations were investigated. IFA, Western blot, and ELISA results showed that the MAbs were specific against JEV NS4B protein. The epitope of the MAbs was further identified using pairs of synthesized overlapping peptides. These MAbs may provide valuable tools for further exploration of the functions of NS4B and the pathogenesis of Japanese encephalitis virus.
日本脑炎(JE)是全球最普遍流行的病毒性脑炎病毒之一。日本脑炎病毒(JEV)NS4B蛋白的功能仍在研究中。在我们的研究中,NS4B在大肠杆菌中表达并通过透析进行纯化。制备了针对NS4B蛋白的两个单克隆抗体克隆(单克隆抗体1B1和1C3)并对其特性进行了研究。免疫荧光分析(IFA)、蛋白质免疫印迹法(Western blot)和酶联免疫吸附测定(ELISA)结果表明,这些单克隆抗体对JEV NS4B蛋白具有特异性。使用合成的重叠肽对进一步鉴定了单克隆抗体的表位。这些单克隆抗体可能为进一步探索NS4B的功能和日本脑炎病毒的发病机制提供有价值的工具。