甲型流感病毒M2蛋白胞外域单克隆抗体的制备及其生物特性  

Preparation and biological characteristics of monoclonal antibodies to extracellular domain of M2 of influenza A virus

作　　者：刘亚辉[1] 沃恩康[1] 杨新燕[1] 徐琦 杨灿 邓冠聪 郭潮潭[1] Liu Yahui;Wo Enkang;Yang Xinyan;Xu Qi;Yang Can;Deng Guancong;Guo Chaotan(Institute of Biological Engineering,Zhejiang A cade my of Medical Sciences,Hangzhou 310013,China)

机构地区：[1]浙江省医学科学院生物工程研究所,杭州310013

出　　处：《国际流行病学传染病学杂志》2018年第6期395-400,共6页International Journal of Epidemiology and Infectious Disease

基　　金：国家自然科学基金(81272551);国家卫生和计划生育委员会科研基金(WKJ-ZJ-1508);浙江省科技计划(2017F10002);浙江省自然科学基金(LGF18C180001、LY18H100005);浙江省医药卫生科技计划(2018KY346).

摘　　要：目的 制备甲型流感病毒高度保守的M2蛋白胞外域(M2e)的单克隆抗体,应用于广谱检测甲型流感病毒。方法 将编码鼠伤寒沙门菌鞭毛蛋白的FljB基因与编码甲型流感病毒M2e的基因进行融合表达,然后纯化蛋白,利用杂交瘤技术制备M2e鼠单克隆抗体。结果 成功表达并纯化了融合蛋白Flagellin-3M2e。制备的2株抗M2e的单克隆抗体3G3和3G4,抗体效价均为105,均能与流感病毒株PR8、Memphis、Aichi结合。3G3可以对MDCK细胞中甲型流感病毒的M2蛋白进行实时定位。结论 成功制备了M2e单克隆抗体,初步具备了临床检测甲型流感的应用基础。Objective To prepare a kind of monoclonal antibodies to the highly conserved extracellular domain of M2 (M2e) of influenza A virus, so as to broadly detect the influenza A virus.Methods The FljB gene coding the flagellin of Salmonella typhimurium and M2e of influenza virus A were fused together, then the fusion protein was purified. The M2e mouse monoclonal antibodies were prepared by hybridoma technique.Results The fusion protein Flagellin-3M2e was successfully expressed and purified. Two types of anti-M2e monoclonal antibodies (3G3 and 3G4) were prepared, and the titer of purified antibodies was 105. The 2 monoclonal antibodies could both react with influenza viruses of PR8, Memphis and Aichi, and 3G3 could be used to locate the M2 protein of influenza A virus in MDCK cells in time.Conclusions Two types of M2e monoclonal antibodies are successfully prepared for clinical detection of influenza A.

关 键 词：流感病毒A型 M2蛋白胞外域 单克隆抗体 鞭毛蛋白 实时定位 

分 类 号：R373.13[医药卫生—病原生物学]