【作者】 靳雯雯； 杨晓红； 邵林； 黄少梅； 陈焕春； 曹胜波；

【Author】 Wenwen Jin,Xiaohong Yang,Lin Shao,Shaomei Huang,Huanchun Chen,Shengbo Cao(College of Veterinary Medicine,Huazhong Agricultural University,Wuhan,Hubei,430070,P.R China)

【机构】 华中农业大学动物医学院；

【摘要】 将人工合成的非洲猪瘟病毒(ASFV)VP73基因在大肠杆菌中进行融合表达。利用纯化的融合蛋白免疫4周龄BALB/c雌性小鼠制备单克隆抗体。通过间接ELISA方法筛选阳性杂交瘤细胞,并最终获得了5株稳定分泌VP73特异单克隆抗体的杂交瘤细胞株,分别命名为1E6、1H3、2H11、3E12、4H5。经抗体亚类鉴定1H3、3E12为IgG1亚类,1E6、2H11、4H5为IgG2b亚类。经Western Blot分析,获得的5株单克隆抗体均能与VP73蛋白发生特异性反应,从而为非洲猪瘟免疫学检测技术的建立奠定了基础。更多还原

【Abstract】 The full-length VP73 gene of ASFV was synthesized,and then VP73 protein was expressed in E.coli efficiently as fusion protein.To prepare the specific monoclonal antibodies against ASFV,GST-VP73 fusion protein expressed in E.coli was used to immunize BALB/c female mice with age of 4 weeks.After detecting and screening by using indirect enzyme linked immunosorbent assay(ELISA),at least five hybridoma cell lines stably secreting VP73-specific monoclonal antibodies were abtained by their strong immunoreactivity,named 1E6、1H3、 2H11、3E12、4H5,respectively.And Subclass identification showed that 1H3、3E12 belonged to IgG1,the others belonged to IgG2b.Then specifity of the McAbs was characterized by western blot and the results showed that these five McAbs were specific to VP73 prokaryotic protein,and they may be used as valuable tools for further establishment of immunological detection technology for ASF.更多还原