论文部分内容阅读
用获得的表达人源性抗ＨＢｓＦａｂ片段的大肠杆菌，发酵表达该抗体的可溶Ｆａｂ片段。以羊抗人Ｆａｂ抗体偶联ＨｉＴｒａｐ柱，用Ａｃｔｉｓｅｐｅｌｕｔｉｏｎｍｅｄｉｕｍ作为洗脱液，在ＦＰＬＣ上纯化。用ＳＤＳＰＡＧＥ及蛋白印迹法分析、鉴定，用固相放兔法检测其活性单位。蛋白印迹及ＥＬＩＳＡ显示转化菌株有抗ＨＢｓＦａｂ片段的表达。经ＦＰＬＣ纯化的抗体Ｆａｂ片段呈单一峰，与抗ＨＢｓ阳性血清相似。纯化后的抗体Ｆａｂ片段达到电泳纯，具有较高的活性。
The obtained soluble Fab fragments of the antibody were fermented using the obtained E. coli expressing the human anti-HBsFab fragment. HiTrap columns were coupled with goat anti-human Fab antibodies and purified on FPLC using Actisepelutionmedium as eluent. SDS PAGE and Western blot analysis, identification, detection of active units by rabbit rabbit solid phase method. Western blotting and ELISA showed that the transformed strains had anti-HBsFab fragment expression. Fab fragments purified by FPLC showed a single peak, similar to anti-HBs positive sera. The purified antibody Fab fragment reached electrophoresis purity, with higher activity.