人源性抗HBsAg单克隆抗体Fab片段的制备及其纯化

目的：制备人源性重组ＨＢｓＡｇ抗体Ｆａｂ片段。方法：采用固相ＨＢｓＡｇ，从已建立的性抗体组成文库中筛选针对ＨＢｓＡｇ的抗体子文库，并转入大肠杆菌中表达。反复冻融细菌获得可溶性Ｆａｂ片段。制备羊抗人ＩｇＧＦａｂ抗体亲和层析柱，纯化Ｆａｂ片段，。ＳＤＳ－ＰＡＧＥ及点印迹法分析纯化后Ｆａｂ片段的纯度及ＨＢｓＡｇ的能力。结果：蛋白印迹显示转化的细菌内有可溶性Ｆａｂ片段的表达。纯化后的Ｆａｂ片段达免疫纯，并

The generation and purification of human monoclonal antibody Fab fragments against HBsAg

Objective: To obtain human monoclonal antibody Fab fragments(Fabs) against HBsAg from combinatorial libraries. Methods: The specific antibody library of anti HBsAg was selected with solid phase HBsAg from established combinatorial library and transformed into E.coli XL1 blue. Souble Fab supernants were obtained through bacterial frozens and thawes. Goat anti human IgG Fab antibody affinity chromatography column was prepared and the Fab supernant was purified. The purity of Fabs was determined with SDS PAGE, and the activity of conjugation with HBsAg was verified with dot blot analysis. Results: The expression of Fabs in bacteria was verified by western blot analysis. Fabs were immunity pure and were able to bind HBsAg. Conclusion: It is probable that human monoclonal antibody Fabs against HBsAg act as clinical reagents.