The Manufacturing Process of Recombinant Factor VIII, Recombinate Edward Gomperts, Roger Lundblad, and Robert Adamson
HE MANVFACTVRING process developed by Genetics Institute, Inc. and Baxter Hyland Division for the production of recombinant factor VIII (rFVIII or rAHF), Recombinate, has been under development since 1984 and was first introduced into cIinical triais in March 1987. 1 The process used to produce the rAHF that is current1y in an advanced stage of elinical tńal is an intricate and complex process that is carefully monitored and controlled to ensure the safety and efficacy of this hemostatic protein in the care of the individual with hemophilia A. This artiele will focus on the major facets of this manufactuńng process incIuding the cell line, rAHF production, rAHF puńfi cation, quality assurance, process validation, and product specifications.
T
CELL L1NE
Chinese Hamster ovary (CHO) cells were chosen as the cellline to produce rFVIII for a number of reasons (Table l). We believed at the beginning ofthe process, and it has proven to be the case over time, that CHO cells synthesize complex recombinant proteins in a manner that is very similar to physiological processing. This has been demonstrated with rFVIII, erythropoietin, tissue plasminogen activator, and with a number of colonystimulating factors. Indeed other investigators have considered this cellline ideal from the point of view of producing both complex glycoproteins and ensuńng maximum equivalence with the native human-deńved proteins.z For example, this cellline does not produce the GAL
HE MANVFACTVRING process developed by Genetics Institute, Inc. and Baxter Hyland Division for the production of recombinant factor VIII (rFVIII or rAHF), Recombinate, has been under development since 1984 and was first introduced into cIinical triais in March 1987. 1 The process used to produce the rAHF that is current1y in an advanced stage of elinical tńal is an intricate and complex process that is carefully monitored and controlled to ensure the safety and efficacy of this hemostatic protein in the care of the individual with hemophilia A. This artiele will focus on the major facets of this manufactuńng process incIuding the cell line, rAHF production, rAHF puńfi cation, quality assurance, process validation, and product specifications.
T
CELL L1NE
Chinese Hamster ovary (CHO) cells were chosen as the cellline to produce rFVIII for a number of reasons (Table l). We believed at the beginning ofthe process, and it has proven to be the case over time, that CHO cells synthesize complex recombinant proteins in a manner that is very similar to physiological processing. This has been demonstrated with rFVIII, erythropoietin, tissue plasminogen activator, and with a number of colonystimulating factors. Indeed other investigators have considered this cellline ideal from the point of view of producing both complex glycoproteins and ensuńng maximum equivalence with the native human-deńved proteins.z For example, this cellline does not produce the GAL
