Abstract
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 γ-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.
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10529_2010_227_MOESM1_ESM.jpg
Supplementary Fig. S1. Light micrographs of Sf9 cells infected with the recombinant baculoviruses at different days post infection. The viability of Sf9 cells starts to drop at day 5 after viral infection. Magnification: 100x. (JPG 1490 kb)
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Masroori, N., Halabian, R., Mohammadipour, M. et al. High-level expression of functional recombinant human coagulation factor VII in insect cells. Biotechnol Lett 32, 803–809 (2010). https://doi.org/10.1007/s10529-010-0227-7
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DOI: https://doi.org/10.1007/s10529-010-0227-7