1. The manufacturing process for B-domain deleted recombinant factor VIII
- Author
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E Lindner-Olsson, B.D Kelley, C Fenge, Timothy S. Charlebois, Å Ljungqvist, A.-L Smeds, J Rosenquist, Mark Leonard, C Ljungqvist, R.K Eriksson, and A Östlin
- Subjects
Genetics ,Cloning ,Factor VIII ,Manufactured Materials ,Chinese hamster ovary cell ,CHO Cells ,Hematology ,Biology ,Human serum albumin ,law.invention ,Column chromatography ,Coagulation ,Biochemistry ,Cell culture ,law ,Cricetinae ,medicine ,Recombinant DNA ,Animals ,Humans ,Cloning, Molecular ,Cell bank ,medicine.drug - Abstract
The development of a cell bank used in the routine manufacturing of a B-domain deleted recombinant coagulation factor VIII (BDDrFVIII) molecule involved stable insertion of the human BDDrFVIII gene into Chinese hamster ovary (CHO) cells, selection of a cell line capable of expressing consistent levels of active BDDrFVIII, and the establishment of a cell bank. The manufacturing process begins with the culturing of CHO cells in large bioreactors. Product synthesis is initiated by altering the cell culture conditions, thereby arresting the cells in a stationary growth phase and inducing elevated expression of BDDrFVIII. Harvested culture medium is concentrated by chromatography and then purified through a series of four column chromatography steps and one solvent-detergent virus inactivation step. By eliminating the presence of human serum albumin in the final formulation, the BDDrFVIII-containing coagulant product meets with a high standard of safety against microbial and viral contamination. Extensive studies have shown that BDDrFVIII is a consistent, highly pure factor VIII for the treatment of patients with hemophilia A.
- Published
- 2001