一种特异性识别斑马鱼卵黄蛋白原的噬菌体展示单链抗体的可溶性表达方案  

Strategy for soluble expression of phage-displayed scFv antibody specific for zebrafish vitellogenin

作　　者：饶瑜[1,2] 钟利桥[1,2] 张凤[1,2] 张晓华[1] 戴和平[1] 

机构地区：[1]中国科学院水生生物研究所淡水生态和生物技术国家重点实验室,武汉430072 [2]中国科学院研究生院,北京100049

出　　处：《生物工程学报》2011年第11期1637-1644,共8页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(No.20977108);淡水生态和生物技术国家重点实验室自主项目(No.2009FBZ08)资助~~

摘　　要：为了实现特异性识别斑马鱼卵黄蛋白原的噬菌体展示单链抗体的可溶性表达,将不能以可溶性蛋白形式表达的、只能以噬菌体展示形式特异性识别斑马鱼卵黄蛋白原的单链抗体F5的基因,克隆到pET 32a载体并转化入大肠杆菌ori DE3中。结果表明,通过诱导表达,可获得可溶性的并且仍特异性识别斑马鱼卵黄蛋白原的单链抗体32a-F5。噬菌体展示单链抗体不能可溶性表达是噬菌体展示技术应用中常见的问题,该方法提供了一种表达可溶性单链抗体的可行性方案。We developed a new method for soluble expression of phage-displayed scFv antibody specific for zebrafish vitellogenin.The scFv antibody F5 could bind zebrafish vitellogenin specifically in phage-displayed form but not soluble form.The gene of scFv antibody F5 was cloned into vector pET 32a and transferred into Escherichia coli ori DE3.With inducible expression,soluble scFv antibody 32a-F5 was obtained successfully and could also specifically bind to zebrafish vitellogenin.The insoluble expression of phage-displayed scFv antibody was a common problem in the practical use of phage display.This study offered a feasible way to express soluble scFv antibodies with biological activity.

关 键 词：噬菌体展示技术 单链抗体 可溶性表达 大肠杆菌 斑马鱼卵黄蛋白原 

分 类 号：Q78[生物学—分子生物学]