摘要
目的：在大肠杆菌中表达抗结肠癌抗体Ｆａｂ片段，比较表达产物和原杂交瘤单克隆抗体的抗原结合特性。方法：以ＤＮＡ重组法构建重组表达质粒，在大肠杆菌中进行表达。对表达产物包涵体进行复性。用ＥＬＩＳＡ法和ＳＡＢＣ法测定复性产物的抗原结合特性。结果：构建了抗结肠癌相关抗原抗体Ｆａｂ片段的重组表达质粒ｐＧＬ４，在大肠杆菌ＪＭ８３中获得表达，表达的Ｆａｂ片段占全菌蛋白的２１％，主要以包涵体形式存在；对包涵体进行分离、变性和复性后得到了有活性的Ｆａｂ分子。结论：复性后的Ｆａｂ具有与原杂交瘤单克隆抗体相同的抗原结合活性。
Objective: To express Fab fragments of the anti colonic cancer antibody and to compare the antigen binding characteristics between the expressing product and the authentic monoclonal antibody. Methods: The expressing vector was constructed by using DNA recombinant methods, transformed into E.coli and induced to express Fab fragments. The characteristic of the product binding to the antigen was detected with ELISA and streptavidin biotin complex. Results: The expressing vector, pGL4, containing the gene encoding Fab fragments was constructed. The recombinant protein was produced mostly in the form of inclusion bodies, the expressing level was about 21% of the total proteins in E.coli JM83. The inclusion bodies were separated, denatured and renatured to achieve native activity. Conclusion: The renatured Fab fragments were demonstrated to have the same antigen binding activity and specificity just as the authentic monoclonal antibody has.
出处
《军事医学科学院院刊》
CSCD
北大核心
1999年第1期13-15,共3页
Bulletin of the Academy of Military Medical Sciences
关键词
FAB片段
大肠杆菌
结肠肿瘤
抗结肠癌抗体
colonic cancer associated antigen
Fab fragment
Escherichia coli
gene expression
colonic neoplasm
