H5N1流感病毒M1蛋白免疫制备的单克隆抗体的研究

目的 制备抗人流感病毒H5N1株M1蛋白的单克隆抗体,为流感的快速诊断和研究提供新的工具.方法 应用在大肠埃希菌中表达的人H5N1亚型禽流感病毒(A/Anhui/1/2005)株M1蛋白,以纯化的表达产物免疫BALB/c小鼠,取脾细胞与sp2/0细胞系作细胞融合后,间接ELISA法筛选阳性的杂交瘤细胞,并应用间接免疫荧光法对抗体的特异性进行鉴定.结果 获得3株能稳定分泌抗禽流感病毒M1抗原的McAb杂交瘤细胞株,交叉反应试验及间接免疫荧光检测表明,三株McAb具有型特异性.结论 用H5N1禽流感病毒M1蛋白免疫制备的单克隆抗体,具有一定的交叉反应性,可用于多种亚型甲型流感病毒的检测.

Study on preparation and identification of monoclonal antibodies immunized with H5N1 influenza virus M1 protein

Objective To prepare monoclonal antibodies specific for M1 protein of H5N1 subtype human influenza virus,this work may provide new tool in rapid diagnosis and study of type A influenza virus.Methods BALB/c mice were immunized with purified recombinant H5N1(A/Anhui/1/2005)/M1 protein expressed in E.coli.Spleen cells of the immunized mice were fused with sp2/0 cells to produce hybridoma cell lines.ELISA was performed to identify the monoclonal antibody against M1 protein of H5N1.Immunofluorescence assay(IFA)were applied to identify the Bpecificity of these antibodies.Results Three hybridoma cell lines steadily secreting anti-H5N1/M1 McAb were obtained,and their cross reactivity was confirmed by cross-reaction test and IfA.Conclusion Monoclonal antibodies immunized with H5N1 subtype influenza virus M1 protein are cross-reactive,which can be used to detect difierent subtype of influenze virus type A.