噬菌体文库与计算机模拟筛选H1N1 流感病毒血凝素结合位点的比较

目的:对H1N1 流感病毒血凝素HA 的抗原表位初步筛选。方法:用噬菌体展示文库对2 株抗体的结合位点进行筛选,将筛选的抗原位点结合HA 序列进行多肽合成,免疫小鼠制备单克隆抗体,通过分子生物学手段对4 株抗体的轻链和重链可变区基因进行调取,通过计算机模拟抗体序列结合的抗原的结构域,推测抗体结合的氨基酸位点。结果:噬菌体文库筛选得出抗体的结合序列富含组氨酸和精氨酸,将针对多肽的单克隆抗体的序列和前期获得的2 株抗HA 单克隆抗体的序列比对分析,发现A1-8 和anti-14p 结合的HA 抗原序列相一致,H1-13 和anti-11p 结合的HA 抗原序列相一致。结论:通过将噬菌体展示文库技术和计算机模拟预测抗原抗体结合,可以有效筛选抗原表位,为流感病毒HA 抗原表位预测方法的完善提供理论基础。

Comparison of hemagglutinin of H1N1 influenza virus binding sites between phage library and computer screening

Objective:To screen the antigenic epitopes of hemagglutinin of H1N1 influenza virus.Methods:The binding sites of the two antibodies with HA were analyzed by phage display library.Polypeptide synthesis based on the selected antigenic sites and HA sequence,monoclonal antibodies prepared by immunization mice with two polypeptide,light chain and heavy chain variable region of four antibodies were obtained by means of molecular cloning.Moreover,the binding domain of antibodies and antigen was predicted by computer simulation.Results:The phage library showed that the binding domain was a sequence of amino acids rich in histidine and arginine.The sequence of HA antigen binding to A1-8 and anti-14p was consistent,and the sequence of HA antigen binding to H1-13 and anti-11p was consistent.Conclusion:Predict antigen-antibody binding sites by the combination of phage display library technology and computer simulation may effectively screen antigen epitopes.This study may provide a theoretical basis for the improvement of influenza antigen HA epitope prediction.