High-level expression of functional recombinant human coagulation factor VII in insect cells.
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.
Masroori
Nasser
N
Research Center, Iranian Blood Transfusion Organization, Tehran, P.O. Box 14665-1157, Iran.
Halabian
Raheleh
R
Mohammadipour
Mahshid
M
Roushandeh
Amaneh Mohammadi
AM
Rouhbakhsh
Mahdi
M
Najafabadi
Ali Jahanian
AJ
Fathabad
Mahdi Edalati
ME
Salimi
Mohamad
M
Shokrgozar
Mohamad Ali
MA
Roudkenar
Mehryar Habibi
MH
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D019735:Carbon-Carbon Ligases / Q000096:biosynthesis / Q000235:genetics; D002460:Cell Line; D005167:Factor VII / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D017422:Sequence Analysis, DNA; D018411:Spodoptera
D011994:Recombinant Proteins; D005167:Factor VII; D019735:Carbon-Carbon Ligases; C020660:glutamyl carboxylase