Research Center, Iranian Blood Transfusion Organization, Tehran, P.O. Box 14665-1157, Iran.
Biotechnol Lett. 2010 Jun;32(6):803-9. doi: 10.1007/s10529-010-0227-7. Epub 2010 Mar 7.
Recombinant coagulation factor VII (FVII) is used as a potential therapeutic intervention in hemophilia patients who produce antibodies against the coagulation factors. Mammalian cell lines provide low levels of expression, however, the Spodoptera frugiperda Sf9 cell line and baculovirus expression system are powerful systems for high-level expression of recombinant proteins, but due to the lack of endogenous vitamin K-dependent carboxylase, expression of functional FVII using this system is impossible. In the present study, we report a simple but versatile method to overcome the defect for high-level expression of the functional recombinant coagulation FVII in Sf9 cells. This method involves simultaneous expression of both human gamma-carboxylase (hGC) and human FVII genes in the host. It may be possible to express other vitamin K-dependent coagulation factors using this method in the future.
重组凝血因子 VII(FVII)被用作产生凝血因子抗体的血友病患者的潜在治疗干预措施。然而,哺乳动物细胞系的表达水平较低,而 Spodoptera frugiperda Sf9 细胞系和杆状病毒表达系统是重组蛋白高水平表达的强大系统,但由于缺乏内源性维生素 K 依赖性羧化酶,使用该系统表达功能性 FVII 是不可能的。在本研究中,我们报告了一种简单但通用的方法,可克服 Sf9 细胞中功能性重组凝血 FVII 的高水平表达缺陷。该方法涉及在宿主中同时表达人γ-羧化酶(hGC)和人 FVII 基因。将来可能使用这种方法表达其他维生素 K 依赖性凝血因子。