title: Mixture of three amino acids as stabilizers replacing albumin in lyophilization of new third generation recombinant factor VIII GreenGene F.
abstract: A formulation with stabilizers replacing albumin was developed for lyophilization of recombinant factor VIII (FVIII), GreenGene F (WHO INN: beroctocog alfa), to achieve stability and eliminate safety issues of blood-derived albumin. L-Arginine (hydrophilic amino acid, positively charged side chain), L-glutamic acid (hydrophilic amino acid, negatively charged side chain), and L-isoleucine (hydrophobic amino acid, nonpolar) were selected as stabilizers, and the mixture of the three amino acids were optimized. The mixture had results comparative with albumin and other commonly used stabilizers showing good preservation of recombinant FVIII during lyophilization, robust stability with consistently high recovery of FVIIII, very low aggregate formation, and good storage stability without alterations in protein characteristics. In vivo test results showed that the efficacy was maintained and had no signs of toxicity. The study demonstrated that the three amino acid mixture acts as a good stabilizer for lyophilization of recombinant FVIII and as a safe excipient.
authors.affiliation: GC-Research Center, Green Cross Corporation, 341 Bojeong-dong, Giheung-gu, Yongin, Korea. shpaik@greencross.com
authors.forename: Sang Hoon
authors.initials: SH
authors.lastname: Paik
authors.forename: Yong Jae
authors.initials: YJ
authors.lastname: Kim
authors.forename: Sang Kyul
authors.initials: SK
authors.lastname: Han
authors.forename: Jean Man
authors.initials: JM
authors.lastname: Kim
authors.forename: Jae Wook
authors.initials: JW
authors.lastname: Huh
authors.forename: Young In
authors.initials: YI
authors.lastname: Park
mesh_terms: D000418:Albumins / Q000737:chemistry; D000596:Amino Acids / Q000737:chemistry* / Q000494:pharmacology; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005079:Excipients / Q000737:chemistry* / Q000494:pharmacology; D005169:Factor VIII / Q000737:chemistry*; D005612:Freeze Drying / Q000379:methods*; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D011136:Polysorbates / Q000737:chemistry; D055550:Protein Stability / Q000187:drug effects; D011994:Recombinant Proteins / Q000737:chemistry*; D013395:Sucrose / Q000737:chemistry
chemical_list: D000418:Albumins; D000596:Amino Acids; D005079:Excipients; D011136:Polysorbates; D011994:Recombinant Proteins; D013395:Sucrose; D005169:Factor VIII