Anna-Lisa Smeds

Development of a Large Scale Process for the Production of Recombinant Truncated Factor VIII in CHO Cells under Cell Growth Arrest Conditions

A large scale biopharmaceutical process has been developed for the production of a genetically engineered variant of coagulation factor VIII. Factor VIII is the coagulation protein that is defective or missing in hemophilia A. Two challenging techniques, ceil growth arrest and perfusion, have been applied to ensure a high production yield. Key development aspects of this process will be presented. A B-domain deleted variant of the factor VIII gene, the factor VIII SQ gene, has been constructed and integrated in Chinese Hamster Ovary CHO DG44 DHFR− cells. The cells are cultivated in serum-free medium which has been developed to optimize the factor VIII SQ production. The cultivation and purification processes have been selected to reduce the deleterious actions of proteases present in the cultivation process. Factor VIII SQ has structural and functional characteristics comparable to full length factor VIII and has been licensed in Europe and USA under the name ReFacto (Genetics Institute).