Central Laboratory, Shaanxi Provincial People's Hospital, Xi'an, 710068, Shaanxi, China.
Shaanxi Provincial Key Laboratory of Infection and Immune Diseases, Xi'an, Shaanxi, China.
Arch Microbiol. 2022 Sep 2;204(9):594. doi: 10.1007/s00203-022-03133-z.
The frequent variation of influenza virus hemagglutinin (HA) antigen is the main cause of influenza pandemic. Therefore, the study of B cell epitopes of HA is of great significance in the prevention and control of influenza virus. In this study, the split vaccine of 2009 H1N1 influenza virus was used as immunogen, and the monoclonal antibodies (mAbs) were prepared by conventional hybridoma fusion and screening techniques. The characteristics of mAbs were identified by ELISA method, Western-blot test and hemagglutination inhibition test (HI). Using the obtained mAbs as a tool, the B cell epitopes of HA were predicted by ELISA blocking test, sandwich ELISA method and computer simulation method. Finally, four mAbs against HA antigen of H1N1 influenza virus were obtained. The results of ELISA and computer prediction showed that there were at least two types of epitopes on HA of influenza virus. The results of this study complemented the existing methods for predicting HA epitopes, and also provided a new method for predicting other pathogenic microorganisms.
流感病毒血凝素 (HA) 抗原的频繁变异是流感大流行的主要原因。因此,研究 HA 的 B 细胞表位对于预防和控制流感病毒具有重要意义。本研究以 2009 年 H1N1 流感病毒的裂疫苗为免疫原,采用常规杂交瘤融合和筛选技术制备单克隆抗体 (mAb)。通过 ELISA 法、Western-blot 试验和血凝抑制试验 (HI) 鉴定 mAb 的特性。利用获得的 mAb 作为工具,通过 ELISA 阻断试验、夹心 ELISA 法和计算机模拟法预测 HA 的 B 细胞表位。最后,获得了针对 H1N1 流感病毒 HA 抗原的 4 株 mAb。ELISA 和计算机预测的结果表明,流感病毒 HA 上至少存在两种类型的表位。本研究补充了现有的 HA 表位预测方法,也为预测其他致病性微生物提供了新方法。