High efficiency recombinant factor VIII production and secretion in CHO cells

Factor VIII is a plasma protein for homeostasis. Haemophilia A, a bleeding disorder, is caused by absence or decreased level of factor VIII. Haemophilia A can be treated by supplementing plasma factor VIII from different sources. In this study, specific human Factor VIII genes that carry variations/mutations for improving protein synthesis and secretion without affecting its function were designed. CHO cell lines were transfected with pOptiVEC plasmids carrying the designed mutant genes. Efficiency of mutant Factor VIII forms was compared against wild type factor VIII in terms of their effect on mRNA synthesis by real time PCR and on protein synthesis by immunoanalysis. According to the data, mutant form of the Factor VIII gene carried by plasmid pC2 (Thr779-Lys1693del) had the highest level of recombinant protein production. Thus, this mutant form can be a good candidate for efficient recombinant Factor VIII production in future.