title: Improved Stability of a Model IgG3 by DoE-Based Evaluation of Buffer Formulations.
abstract: Formulating appropriate storage conditions for biopharmaceutical proteins is essential for ensuring their stability and thereby their purity, potency, and safety over their shelf-life. Using a model murine IgG3 produced in a bioreactor system, multiple formulation compositions were systematically explored in a DoE design to optimize the stability of a challenging antibody formulation worst case. The stability of the antibody in each buffer formulation was assessed by UV/VIS absorbance at 280 nm and 410 nm and size exclusion high performance liquid chromatography (SEC) to determine overall solubility, opalescence, and aggregate formation, respectively. Upon preliminary testing, acetate was eliminated as a potential storage buffer due to significant visible precipitate formation. An additional 2(4) full factorial DoE was performed that combined the stabilizing effect of arginine with the buffering capacity of histidine. From this final DoE, an optimized formulation of 200 mM arginine, 50 mM histidine, and 100 mM NaCl at a pH of 6.5 was identified to substantially improve stability under long-term storage conditions and after multiple freeze/thaw cycles. Thus, our data highlights the power of DoE based formulation screening approaches even for challenging monoclonal antibody molecules.
authors.affiliation: Division II, Office of Biotechnology Products, OPQ, CDER, FDA, Silver Spring, MD 20903, USA.
authors.forename: Brittany K
authors.initials: BK
authors.lastname: Chavez
authors.affiliation: Division of Product Quality Research, Office of Testing and Research, OPQ, CDER, FDA, Silver Spring, MD 20903, USA.
authors.forename: Cyrus D
authors.initials: CD
authors.lastname: Agarabi
authors.affiliation: Division II, Office of Biotechnology Products, OPQ, CDER, FDA, Silver Spring, MD 20903, USA.
authors.forename: Erik K
authors.initials: EK
authors.lastname: Read
authors.affiliation: Division of Pharmaceutical Analysis, Office of Testing and Research, OPQ, CDER, FDA, Silver Spring, MD 20903, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Boyne
authors.affiliation: Division of Product Quality Research, Office of Testing and Research, OPQ, CDER, FDA, Silver Spring, MD 20903, USA.
authors.forename: Mansoor A
authors.initials: MA
authors.lastname: Khan
authors.affiliation: Division II, Office of Biotechnology Products, OPQ, CDER, FDA, Silver Spring, MD 20903, USA.
authors.forename: Kurt A
authors.initials: KA
authors.lastname: Brorson
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D000917:Antibody Formation; D002021:Buffers; D002851:Chromatography, High Pressure Liquid; D005615:Freezing; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology; D051379:Mice; D055550:Protein Stability
chemical_list: D000911:Antibodies, Monoclonal; D002021:Buffers; D007074:Immunoglobulin G