抗甲型肝炎病毒抗体Fab在大肠杆菌中的表达

目的 在大肠杆菌中高效表达人源抗HAV抗体Fab，为进一步研究该抗体的功能奠定基础。方法 依次将抗HAV抗体的Fd及轻链基因克隆到表达载体pASK84的不同信号序列下，构建成Fab表达质粒，在大肠杆菌JM83中进行表达，并对表达产物进行复性。用SDS－PAGE、western blot和ELISA法检测Fab的表达及其与甲肝病毒抗原的结合活性。结果 构建了抗HAV Fab的重组表达质粒p84Fab，在大肠杆菌JM83中获得表达，表达的Fab占全菌的25％，主要以包涵体形式存在，复性后具有抗原结合活性；培养基、菌体可溶性蛋白部分也能检测到有抗原结合活性的Fab。结论 抗HAV抗体Fab在大肠杆菌中得到了表达，并具有良好的抗原结合活性。

Expression of human anti-HAV Fab in E.coli JM83

Objective To construct expression plasmids for human anti-HAV Fab which was screened from phage display antibody library and express efficently the Fab in E.coli. Methods Expression vector p84Fab was constructed by DNA recombination technique, then transformed into E.coli JM83. SDS-PAGE and western blot was used to detect the expression of Fab,ELISA revealed the binding activity to HAV antigen. Results Expression vector was successfully constructed, and anti-HAV Fab was produced mostly in the form of inclusion bo-dies, the expressing level was about 25% of the total proteins in JM83. The inclusion bodies were separated, denatured and restored succes-sfully. Conclusionanti-HAV Fab were produced in E.coli efficiently and exhibited the binding activity to HAV antigen.