H1N1流感病毒HA蛋白抗原表位预测方法研究

Study on antigenic epitope prediction methods of HA protein of H1N1 influenza virus

目的对H1N1流感病毒血凝素（hemagglutinin，HA）蛋白抗原表位预测方法进行研究。方法用H1N1流感病毒裂解疫苗常规法免疫BALB／c小鼠，取脾细胞与骨髓瘤细胞（SP2／O）融合、克隆化筛选，获得特异性单克隆抗体（monoclonal antibody，mAb），辣根过氧化物酶（horseradish peroxidase，HRP）标记各株mAb，ELISA阻断试验检测各mAb相互阻断结果；克隆各mAb的轻链和重链可变区基因，并用计算机模拟预测各mAb结合HA抗原表位的氨基酸位点。结果获得稳定分泌抗H1N1流感病毒HA蛋白的杂交瘤细胞3株；ELISA阻断试验将3株mAbs分为2类；计算机模拟预测将3株mAbs也分为2类。结论ELISA阻断试验和计算机模拟预测在解析H1N1流感病毒HA蛋白抗原表位时可以相互佐证。

Objective To investigate the antigenic epitope prediction method of hemagglutinin （HA） protein of influenza A （H1N1） virus. Methods BALB/c mice were conventionally immunized with influenza H1N1 vaccine. The splenocytes from the immunized mouse were fused with SP2/O mouse myeloma cell line, and then the antigen-specific monoclonal antibodies （mAbs） were obtained by screening hybridoma supernatants. ELISA blocking test was used to detect the blocking result of each monoclonal antibody, which was labeled by horseradish peroxidase （HRP）. The light and heavy chain variable region genes of each antibody were cloned, the amino acid sites of the antibody-binding HA antigen epitope were predicted by computer simulation. Results Three hybridoma cell lines of stable secreting anti-H1N1 influenza virus HA protein were obtained. Three mAbs were divided into two categories by ELISA blocking tests, which were divided into two categories according to preliminary results of computer simulation. Conclusions ELISA blocking test and computer simulation prediction can prove each other in predication of the antigenic epitopes of HA protein of H1N1 influenza virus.