Evaluation of different concentrations of vitamin K in the production of recombinant coagulation factor IX

Introduction : Hemophilia B is an of X- linked recessive blood clotting disorders caused by a mutation in the factor IX gene, leading to a deficiency of factor IX. It is the second-most common form of hemophilia, rarer than hemophilia A, which occurs in 1/30,000 males. Treatment or prevention of bleeding episodes in hemophilia B patients is based on the injection of coagulation factor IX (replacement therapy) concentrated from pooled human plasma or from recombinant preparations. Methods: To evaluate effect of vitamin K in activation of factor IX, in this study, human factor IX cDNA was transfected into Drosophila S2 cells using an expression vector (pMT / V5). Experiments were carried out at 5 concentrations of vitamin K (0.1 - 1 – 5 -10 μg / ml) and without vitamin K (0) at three-day intervals and it was determined that the cells at the concentration of 5 µg/ml had most expressed. Also, expression comparisons for different concentrations of vitamin K in periods of three days indicated that in all concentrations, the highest expression of Factor IX was obtained in 72 hours. Results: In this study, we showed that the concentration of vitamin K could have an effect on the production rate of active factor IX and it is noted that S2 cells in the presence of vitamin K can done undergo changes in gamma - carboxylation and produce biologically active protein.