4株埃博拉病毒核蛋白特异性单克隆抗体抗原结合表位研究  

Study on the epitopes of four monoclonal antibodies specific to ebola virus nucleoprotein

作　　者：孙丽娜[1] 周荣苹 刘洋[1] 芜为[1] 李川[1] 仇佩虹[2] 李德新[1] 梁米芳[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206 [2]温州医科大学,325035

出　　处：《中华实验和临床病毒学杂志》2016年第2期138-140,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的 分析鼠埃博拉病毒核蛋白（NP）特异性单克隆抗体抗原结合表位,了解其免疫学特性.方法 经生物信息学分析后,将NP分段重组表达,并分别以之为抗原,通过免疫印迹和竞争ELISA的方法对4株埃博拉病毒核蛋白特异性单抗的抗原结合表位进行分析.结果 经生物信息学表位分析后将埃博拉病毒全长核蛋白分为rNP1-418,rNP419-639和rNP640-739三段进行重组制备,免疫印迹法分析表明4株鼠单抗均识别线性表位,结合位点位于C端100 aa.通过对C端100 aa即rNP640-739 N端连续截短20 aa的方法进行鉴定,4株鼠单抗抗原表位位于埃博拉核蛋白C末端20 aa,但其中2株单抗无明显竞争.结论 明确了具有不同结合位点的埃博拉核蛋白特异性单抗,为免疫学快速检测试剂的研究奠定基础.Objective To analysis the epitope of Ebola virus （EBOV） nucleoprotein （NP） and understand its immunological properties.Methods Firstly four mouse mAbs recognizing the different epitopes of Ebola virus NP were classified by competition ELISA.The full-length NP and the C-terminal were truncated and expressed in E.coli.according to epitope prediction by software and some published papers.Epitope mapping of mAbs to a panel of the truncated rNP were identified in WB analysis.Results After epitope prediction,the full-length NP were divided into rNP1-418,rNP419-639 and rNP640-739.WB analysis indicated that mAbs all recognized the linear epitopes,which were limited in the C-terminal 100aa （rNP640-739）.The truncated N proteins with N-terminal of rNP640-739 series 20aa deletions reacted with all mAbs.The epitopes recognized by anti-NP mAbs were defined near the C-terminal 20aa of EBOV NP.Conclusion Analysis of antigenic epitopes of EBOV NP by specific mAbs as a probe lay the foundation for the study of immunology diagnostic reagents.

关 键 词：埃博拉病毒 核蛋白 抗原表位 

分 类 号：R392[医药卫生—免疫学]