运用单克隆抗体分析流感病毒H1亚型HA蛋白的抗原表位

摘要

目的 对甲型流感病毒H1亚型血凝素蛋白的抗原表位进行分析.方法 以前期获得的97株抗H1亚型HA抗原的单克隆抗体为研究工具,用间接ELISA方法对不同亚型的血凝素抗原进行反应,将H1亚型HA蛋白的抗原表位进行粗分类,用ELISA阻断试验对抗原表位进行细分类.选取两类抗原表位的抗体,通过行计算机模拟分析抗体与抗原结合的结构域,推测抗体结合的具体氨基酸位点.结果 根据抗体的交叉反应性不同,将流感病毒Ⅱ1亚型的HA的抗原表位分为8类；根据ELISA阻断试验的初步结果,H1亚型HA抗原的抗原表位又被细分为15类,通过计算机模拟分析预测的抗体与流感病毒HA抗原结合的氨基酸位点结果与ELISA阻断试验结果相吻合.结论 证明流感病毒H1N1亚型HA抗原上至少含有15个抗原表位,且通过ELISA阻断试验和计算机模拟分析得到了相互印证,为流感病毒血凝素HA抗原表位预测方法的完善及通用疫苗的研制提供了实验数据.%Objective To analyze the hcmagglutinin (HA) antigenic epitopes of HI influenza virus. Methods First, we used 97 monoclonal antibodies to HI influenza virus HA antigen and classified antigenic category by means of indirect ELISA. Then, an epitope classification was made through blocking ELJSA test. Finally, the antibodies and antigen binding domain was analyzed by computer modeling to predict the possible binding sites. Results HA epitopes of influenza virus H1 subtype were classified into eight categories according to antibody cross-reactivity. Then HA antigenic epitopes of H1 subtype were divided into fifteen categories according to the preliminary results of ELISA blocking experiments. The results of computer simulation were consistent with the data of blocking ELISA test. Conclusion The paper classified HA antigen of H1N1 subtype of influenza virus into at least fifteen epitopes, and the results of ELISA blocking experiments and computer simulations confirmed with each other. This study provides available experimental data for improving prediction methods of HA epitope of Hl-type influenza virus and the development of universal vaccine.