鸭坦布苏病毒E蛋白单克隆抗体的制备及鉴定

Preparation and identification of monoclonal antibody to E protein of duck Tembusu virus

用原核表达并纯化的鸭坦布苏病毒(duck Tembusu virus,DTMUV)E蛋白为抗原免疫BALB/c小鼠,应用淋巴细胞杂交瘤技术,筛选到1株分泌针对DTMUV的E蛋白的单抗融合细胞,并命名为LY12。Western blot和间接免疫荧光试验(IFA)分析表明,所获得细胞分泌的抗体只与DTMUV的E蛋白发生特异性反应,与日本脑炎病毒(JEV)和西尼罗病毒(WNV)无交叉反应。进一步鉴定该单抗的抗体亚类为IgG1,轻链为κ链。抗E蛋白单克隆抗体的制备为DTMUV的诊断和相关基础研究奠定了基础。

To prepare a monoclonal antibody （MAb） to duck Tembusu virus, （DTMUV）, BALB/c mice were immunized by the purified protein E of DTMUV, and spleens were used for fusion with SP2/0. One hybridoma secreting MAb were selected by indirect enzyme-linked immunosorbent assay （ELISA）. Western blot and indirect immunofluoreseence assay （IFA） test analysis showed the MAb reacted with DT-MUV, but had no cross reaction to Japanese encephalitis virus （JEV） and West Nile virus （WNV）. The immunoglobulin subtype of the MAh was IgG1. The MAb could be used to establish the monitoring and differential diagnosis methods for DTMUV infection.