鸭坦布苏病毒E蛋白线性化B细胞抗原表位的鉴定

摘要

This experiment was conducted to identify B cell epitope of E protein of duck Tembusu virus ( DTMUV) . In this study, a linear B cell epitope 385LVGSGKGQI393(EP385) for DTMUV E protein were identified by truncated expressing the third domain of E protein (Domain III, DI I), which is recognized by monoclonal antibody BD 10 pre-pared by the purified DTMUV YY 5 strain as antigen , and then by scanning the peptide library which containing 15 o-verlapping segment of truncated and fused expressed D I I with BD10.Then, the immunogenicity of EP385 was veri-fied by fusion expressed in E.coli and immunized mice to prepare a polyclonal antibody which was confirmed to be capable of recognizing viral E protein .The identification of B cell epitope for the E protein of DTMUV may lead to the preparation of a DTMUV peptide vaccine and the development of specific serological diagnostic methods .%为鉴定鸭坦布苏病毒(duck Tembusu virus,DTMUV)的B细胞抗原表位,以纯化的DTMUV YY5株为抗原制备了2株单克隆抗体AE4和BD10,研究证实BD10为线性化表位,其结合的抗原表位位于E蛋白的第三结构域(DomainⅢ,DⅢ).将DTMUV E蛋白DⅢ结构域基因截短为具有末端相互重叠的15段,与MBP融合表达后以单克隆抗体BD10进行肽库扫描,结果筛选出DTMUV一个B细胞线性表位385 LVGSGKGQI393(EP385).该表位原核融合表达产物免疫小鼠制备的抗体,能够和DTMUV E蛋白反应,表明EP385表位具有良好的免疫原性,可为DTMUV多肽疫苗的研制及特异血清学诊断方法的开发奠定基础.