title: Manufacturing process used to produce long-acting recombinant factor VIII Fc fusion protein.
abstract: Recombinant factor VIII Fc fusion protein (rFVIIIFc) is a long-acting coagulation factor approved for the treatment of hemophilia A. Here, the rFVIIIFc manufacturing process and results of studies evaluating product quality and the capacity of the process to remove potential impurities and viruses are described. This manufacturing process utilized readily transferable and scalable unit operations and employed multi-step purification and viral clearance processing, including a novel affinity chromatography adsorbent and a 15 nm pore size virus removal nanofilter. A cell line derived from human embryonic kidney (HEK) 293H cells was used to produce rFVIIIFc. Validation studies evaluated identity, purity, activity, and safety. Process-related impurity clearance and viral clearance spiking studies demonstrate robust and reproducible removal of impurities and viruses, with total viral clearance >8-15 log10 for four model viruses (xenotropic murine leukemia virus, mice minute virus, reovirus type 3, and suid herpes virus 1). Terminal galactose-α-1,3-galactose and N-glycolylneuraminic acid, two non-human glycans, were undetectable in rFVIIIFc. Biochemical and in vitro biological analyses confirmed the purity, activity, and consistency of rFVIIIFc. In conclusion, this manufacturing process produces a highly pure product free of viruses, impurities, and non-human glycan structures, with scale capabilities to ensure a consistent and adequate supply of rFVIIIFc.
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: justin.mccue@biogen.com.
authors.forename: Justin
authors.initials: J
authors.lastname: McCue
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: rashmi.kshirsagar@biogen.com.
authors.forename: Rashmi
authors.initials: R
authors.lastname: Kshirsagar
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: keith.selvitelli@biogen.com.
authors.forename: Keith
authors.initials: K
authors.lastname: Selvitelli
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: qi.lu@biogen.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Lu
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: mingxuan.zhang@biogen.com.
authors.forename: Mingxuan
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: baisong.mei@biogen.com.
authors.forename: Baisong
authors.initials: B
authors.lastname: Mei
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: robert.peters@biogen.com.
authors.forename: Robert
authors.initials: R
authors.lastname: Peters
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: gfp555@gmail.com.
authors.forename: Glenn F
authors.initials: GF
authors.lastname: Pierce
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: jennifer.dumont@biogen.com.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Dumont
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: stephen.raso@biogen.com.
authors.forename: Stephen
authors.initials: S
authors.lastname: Raso
authors.affiliation: Biogen, 14 Cambridge Center, Cambridge, MA 02142, USA. Electronic address: heidi.reichert-robes@biogen.com.
authors.forename: Heidi
authors.initials: H
authors.lastname: Reichert
mesh_terms: D003692:Delayed-Action Preparations; D005169:Factor VIII / Q000096:biosynthesis* / Q000302:isolation & purification / Q000627:therapeutic use; D057809:HEK293 Cells; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000627:therapeutic use
chemical_list: D003692:Delayed-Action Preparations; D011993:Recombinant Fusion Proteins; D005169:Factor VIII
keywords: HEK 293H; Hemophilia A; Human cell line; Manufacturing; Recombinant; rFVIIIFc