应用分子伴侣探索e23sFv/His融合蛋白的可溶性表达  

Application of molecular chaperones to soluble expression of e23sFv/His fusion proteins

作　　者：常镜 欧阳清[1] 杜晓[1] 杨安钢[1] 赵晶[1] 阎博[1] 

机构地区：[1]第四军医大学生物化学与分子生物学教研室,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2015年第9期1205-1210,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81372459)

摘　　要：目的探讨利用分子伴侣实现e23sFv/His融合蛋白可溶性表达的可行性。方法将分子伴侣质粒pGro7、pKJE7与e23sFv原核表达载体共转化BL21(DE3)感受态细胞。16℃诱导融合蛋白可溶性表达,比较可溶性表达产物相对于常规包涵体不溶形式表达产物在产量和抗原结合活性方面的差异。结果可溶性表达的e23sFv/His产量较小,纯化得率低,其抗原结合活性没有明显优于包涵体表达纯化产物。结论应用pGro7、pKJE7有助于e23sFv/His融合蛋白的可溶性表达,但其表达和纯化得率较低。Objective To explore the feasibility of applying molecular chaperones to the soluble expression of e23sFv/His fusion proteins. Methods The molecular chaperone plasmid pGro7 or pKJE7 was transformed into BL21（ DE3） competent cells together with the prokaryotic expression vector harboring His-tagged e23sFv. The soluble expression of e23sFv / His proteins was induced at 16℃. The yield and antigen-binding activity of the soluble products were compared with those of the insoluble products conventionally purified from inclusion bodies. Results Both the overall yield and the purification ratio of soluble e23sFv / His proteins were relatively lower. The binding affinity of the soluble products to immobilized HER2 was not superior to that of the insoluble products from inclusion bodies. Conclusion The molecular chaperone plasmids pGro7 and pKJE7 partially facilitate the soluble expression of e23sFv / His proteins,but both the yield and the purification ratio are stil limited.

关 键 词：e23sFv 可溶性表达 分子伴侣 

分 类 号：R392-11[医药卫生—免疫学] R392-33[医药卫生—基础医学]