人源抗乙型肝炎病毒基因工程全抗体的研制

Development of recombinant human-derived IgG antibody against HBsAg

目的 研制人源化抗乙型肝炎表面抗原 (HBsAg)的IgG全抗体。方法 采用噬菌体抗体库技术 ,从乙肝疫苗免疫后健康人外周血淋巴细胞中提取抗体基因建立抗体库 ,用纯化的HBsAg筛选Fab抗体 ;再将获得的抗体轻、重链构建到杆状病毒表达载体pAc κ Fc中 ,转染SF9细胞表达IgG全抗体并进行纯化。免疫荧光检测抗体的表达 ,竞争抑制ELISA检测其与抗原的结合活性及特异性。结果 获得了一株抗乙肝表面抗原的Fab抗体 ;在转染后的SF9培养上清中收获到了IgG抗体 ,竞争ELISA证实该抗体针对HBV表面抗原的特异性抗体。结论 本实验得到了一株人源抗HBsAg的IgG抗体 ,并进行了纯化。

Objective To investigate preparation of human-derived anti-HBs IgG antibody. Methods mRNA was extracted from lymphocytes of a volunteer with high level antibody to HBsAg. Fab phage display library was constructed. Purified HBsAg was used to screen the Fab. Eukaryotic expression vector was constructed and transfected into sf9 cells,then the supernatant was purified. The expression of anti-HBs human IgG was tested with blocking ELISA and immunofluorescent test. Results A whole human anti-HBs IgG molecule was obtained,the antibody reacts with HBsAg “a” determinant. Conclusion It can be concluded preliminarily that this antibody has neutralizing characteristic against HBsAg.