人源性TAT-Fab抗体的表达优化及生物活性测定

Optimization of human TAT-Fab antibody expression and its bioactivity assay.

为了提高抗体的表达水平,获得有保护作用的人源性破伤风抗毒素Fab(TAT-Fab)抗体,对人源性TAT-Fab抗体在大肠杆菌中的表达条件进行优化并测定其体内外生物学活性.ELISA显示所表达的TAT-Fab抗体可特异性结合破伤风外毒素,且优化后的表达水平较优化前提高近5倍;表达产物经超滤分离浓缩后,Western-blot显示在分子量约50 kDa处有一TAT-Fab抗体的特异性表达条带;小鼠保护实验显示其可特异性中和破伤风外毒素,明显延长小鼠的生存时间.以上结果表明所制备的人源性TAT-Fab抗体在体内外均具有中和破伤风外毒素的效应,为人源性破伤风抗毒素的研制和生产提供了实验依据.

In order to establish high effective expression system for human TAT-Fab antibody with protective activity, the induction conditions for the expression of TAT-Fab antibody in E. coli were optimized, and the bioactivity of antibody was assayed in vitro and in vivo. ELISA showed that human TAT-Fab antibody expressed in E. coli possessed binding activity with tetanus toxin, and the expression level increased to nearly 5 times as compared with that before optimization. The expression products were ultra-filtered and analyzed by Western-blot, a specific band of TAT-Fab antibody, about 50 kDa, was detected. This antibody could neutralized tetanus toxin specifically and prolonged survival time of the mouse in vivo. The results indicate that human TAT-Fab antibody has the effect of neutralizing tetanus toxin in vitro and in vivo, which will provide experimental data for the production of human TAT antibody in the future.