H1N1流感病毒血凝素B细胞抗原表位的鉴定

目的利用计算机模拟和ELISA阻断实验研究流感病毒血凝素(HA)B细胞抗原表位,建立病原微生物表位检测新方法。方法以2009年H1N1流感病毒裂解疫苗作为免疫原,采用常规杂交瘤融合、筛选技术制备单克隆抗体(mAb),应用ELISA、血凝抑制试验(HI)及Western blot法鉴定获得mAb的特性。以获得的mAb为工具,联合应用ELISA阻断实验和计算机模拟方法预测H1N1流感病毒的B细胞抗原表位。结果获得4株抗H1N1流感病毒HA抗原的mAb,通过ELISA阻断实验将HA的B细胞抗原表位分为两类,通过计算机模拟预测发现4株抗体能与HA上的两类表位相结合。结论计算机模拟和ELISA阻断实验的结果一致,建立了用于预测其他病原微生物表位的新方法。

Identification of B cell epitopes on hemagglutinin of H1N1 influenza virus

Objective To study B cell epitopes of the influenza virus hemagglutinin(HA) by computer simulation and ELISA blocking experiment, and to establish a new method for pathogen microbial epitope detection. Methods The 2009 H1N1 influenza virus inactivated vaccine was used as the immunogen, and the monoclonal antibodies(mAbs) were prepared by conventional hybridoma fusion and screening techniques. The characteristics of mAbs were identified by ELISA, hemagglutination inhibition(HI) and Western blot analysis. The obtained mAbs were used as a tool to predict the epitope of H1N1 influenza virus by ELISA blocking experiment combined with computer simulation method. Results Four mAbs against the HA antigen on H1N1 influenza virus were obtained, and HA epitopes were classified into two types by ELISA blocking experiment. The computer simulation revealed that the four antibodies could bind to two epitopes on HA. Conclusion The results of computer simulation and ELISA blocking experiment are consistent, and a new method for predicting the epitopes of other pathogenic microorganisms had been established.