【摘要】 本研究旨在制备非洲猪瘟病毒(ASFV)S273R蛋白的特异性单克隆抗体。本研究以原核表达的非洲猪瘟病毒重组S273R蛋白免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞进行融合获得杂交瘤细胞。结果显示:基于纯化的S273R蛋白建立的间接ELISA方法对杂交瘤细胞进行筛选和亚克隆,获得了4株可稳定分泌抗ASFV S273R蛋白单克隆抗体的杂交瘤细胞株,并通过免疫印迹(Western blot)和免疫荧光试验(immunofluorescence assay, IFA)测定了其免疫特性。4株单克隆抗体亚类均为IgG1型,轻链均为κ链。本研究获得的非洲猪瘟病毒S373R蛋白单克隆抗体可为进一步研究ASFV pS273R的生物学功能及基因缺失毒株的鉴别诊断和疫苗开发提供技术储备。更多还原

【Abstract】 The objective of this study was to prepare monoclonal antibodies against the S273 R protein of African swine fever virus(ASFV). BALB/c mice were immunized with purified recombinant S273 R protein expressed by prokaryotic expression system and hybridoma cells were obtained by fusion of spleen cells and myeloma cells. An indirect ELISA based on the purified S273 R protein was developed. Four hybridoma cell lines stably secreting MAbs against the S273 R protein were harvested after screening and subcloning, and their sensitivity and specificity were detected by ELISA, Western blot, and immunofluorescence assay(IFA). Four MAbs heavy chains belong to IgG1, and light chains were κ. This research provided technical support for further study of biological functions of ASFV pS273 R, as well as its gene-deletion based vaccine development.更多还原