鸭坦布苏病毒E蛋白线性化B细胞抗原表位的鉴定

Identification of linear B cell epitope for E protein of duck Tembusu virus

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,D_Ⅲ)。将DTMUV E蛋白D_Ⅲ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)LVGSGKGQI_(393)(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。

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_(385) LVGSGKGQI_(393)( EP385) for DTMUV E protein were identified by truncated expressing the third domain of E protein( Domain III,D_Ⅲ ),which is recognized by monoclonal antibody BD10 prepared by the purified DTMUV YY5 strain as antigen,and then by scanning the peptide library which containing 15 overlapping segment of truncated and fused expressed D_Ⅲ with BD10. Then,the immunogenicity of EP385 was verified 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.