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

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.