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鼠抗SARS病毒X4蛋白单克隆抗体的制备及鉴定
引用本文:罗昕,张婷,马大龙,陈英玉.鼠抗SARS病毒X4蛋白单克隆抗体的制备及鉴定[J].北京大学学报(医学版),2005,37(5):485-488.
作者姓名:罗昕  张婷  马大龙  陈英玉
作者单位:北京大学人类疾病基因研究中心,北京,100083
摘    要:目的:制备SARS病毒X4蛋白的单克隆抗体,探讨X4蛋白的结构和功能,以及临床应用的价值.方法:以重组SARS病毒X4蛋白为免疫原,免疫Balb/c小鼠,取免疫小鼠的脾细胞和同系小鼠的骨髓瘤细胞进行常规融合, 通过间接ELISA的筛选和有限稀释克隆化,获得鼠抗SARS病毒X4蛋白单克隆抗体的杂交瘤细胞株,通过ELISA,Western blot和免疫荧光实验等方法鉴定其特性.结果:成功地建立了3株稳定分泌抗X4蛋白的单克隆抗体杂交瘤细胞株,分别命名为8A5,8H6和4A2.3株单克隆抗体的免疫球蛋白亚类均为IgG2a.通过ELISA, Western blot等方法的鉴定,抗X4蛋白的单克隆抗体能与X4蛋白发生特异性反应.免疫荧光实验的结果表明,X4蛋白的单克隆抗体能与SARS-病毒感染的Vero E6 细胞发生特异性结合.结论:获得了特异性识别SARS病毒X4蛋白的单克隆抗体,为SARS病毒X4蛋白的生物学功能研究奠定了基础.

关 键 词:严重急性呼吸综合征  SARS病毒  抗体  单克隆  SARS  病毒  免疫球蛋白亚类  单克隆抗体  protein  monoclonal  antibodies  identification  功能研究  生物学  特异性识别  特异性结合  细胞发生  Vero  感染  特异性反应  体能  命名  稳定  结果  特性
文章编号:1671-167X(2005)05-0485-04
修稿时间:2005年2月23日

Preparation and identification of monoclonal antibodies against SARS-CoV putative protein X4
LUO Xin,ZHANG Ting,MA Da-long,CHEN Ying-yu.Preparation and identification of monoclonal antibodies against SARS-CoV putative protein X4[J].Journal of Peking University:Health Sciences,2005,37(5):485-488.
Authors:LUO Xin  ZHANG Ting  MA Da-long  CHEN Ying-yu
Institution:Center for Human Disease Genomics, Peking University, Beijing 100083, China.
Abstract:Objective: To obtain monoclonal antibodies against putative protein X4 of SARS-CoV for further study of the structure and function of protein X4. Methods: Balb/c mice were immunized with recombinant Protein X4, hybridoma cell lines secreting monoclonal antibodies against protein X4 were screened by regular cell fusion and subcloning approach. The specificities of these monoclonal antibodies were determined by Western blotting and Immunofluorescecence assay. Results: Three hybridoma cell lines (8A5, 8H6 and 4A2) stable in secreting specific monoclonal antibodies were successfully obtained. They could bind specifically to protein X4 proved by Western blotting. All of them belonged to the IgG2a isotype proved by antigen mediated ELISA. Indirect Immunofluorescence assay indicated that they could specifically bind to protein X4 expressed in SARS-CoV infected Vero E6 cells. Conclusion: Monoclonal antibodies of high specificity against protein X4 have been successfully prepared, which laid the foundation for the further study of protein X4.
Keywords:Severe acute respiratory syndrome  SARS virus  Antibodies monoclonal
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