高效表达活性重组组织因子

issue factor (TF) is an important glycoprotein in the human body. When blood vessels or tissues are damaged, TF initiates the blood coagulation reaction by activating the extrinsic coagulation pathway. In recent years, the market demand for TF has increased rapidly due to its key role in preoperative coagulation testing. Currently, although recombinant TF has been successfully expressed in some expression systems, large-scale production of active TF is still a challenge because the activity of TF is affected by protein post-translational modifications.N-glycosylation modification, as one of the important ways of post-translational modification of proteins, plays important functions in protein folding, transport and activity. In this study, recombinant human TF was expressed in two expression systems, prokaryotic Escherichia coli and eukaryotic HEK293 cells, and their activities were compared. The TF activity derived from HEK293 cells was higher than that derived from E. coli. Follow-up researches demonstrated the importance of N-glycosylation modification for TF stability and activity. In addition, knocking out the E3 ubiquitin ligase HRD1 in the endoplasmic reticulum-related degradation pathway can improve TF stability without affecting its activity. The research provides a new idea for the large-scale production of TF.