title: Examination of thermal unfolding and aggregation profiles of a series of developable therapeutic monoclonal antibodies.
abstract: Screening for pharmaceutically viable stability from measurements of thermally induced protein unfolding and short-term accelerated stress underpins much molecule design, selection, and formulation in the pharmaceutical biotechnology industry. However, the interrelationships among intrinsic protein conformational stability, thermal denaturation, and pharmaceutical stability are complex. There are few publications in which predictions from thermal unfolding-based screening methods are examined together with pharmaceutically relevant long-term storage stability performance. We have studied eight developable therapeutic IgG molecules under solution conditions optimized for large-scale commercial production and delivery. Thermal unfolding profiles were characterized by differential scanning calorimetry (DSC) and intrinsic fluorescence recorded simultaneously with static light scattering (SLS). These molecules exhibit a variety of thermal unfolding profiles under common reference buffer conditions and under individually optimized formulation conditions. Aggregation profiles by SE-HPLC and bioactivity upon long-term storage at 5, 25, and 40 °C establish that IgG molecules possessing a relatively wide range of conformational stabilities and thermal unfolding profiles can be formulated to achieve pharmaceutically stable drug products. Our data suggest that a formulation design strategy that increases the thermal unfolding temperature of the Fab transition may be a better general approach to improving pharmaceutical storage stability than one focused on increasing Tonset or Tm of the first unfolding transition.
authors: Protein Pharmaceutical Development, Biogen Idec, 14 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors: Mark L
authors: ML
authors: Brader
authors: Protein Pharmaceutical Development, Biogen Idec, 14 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors: Tia
authors: T
authors: Estey
authors: Protein Pharmaceutical Development, Biogen Idec, 14 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors: Shujun
authors: S
authors: Bai
authors: Protein Pharmaceutical Development, Biogen Idec, 14 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors: Roy W
authors: RW
authors: Alston
authors: Protein Pharmaceutical Development, Biogen Idec, 14 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors: Karin K
authors: KK
authors: Lucas
authors: Protein Pharmaceutical Development, Biogen Idec, 14 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors: Steven
authors: S
authors: Lantz
authors: Protein Pharmaceutical Development, Biogen Idec, 14 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors: Pavel
authors: P
authors: Landsman
authors: Protein Pharmaceutical Development, Biogen Idec, 14 Cambridge Center, Cambridge, Massachusetts 02142, United States.
authors: Kevin M
authors: KM
authors: Maloney
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002152:Calorimetry, Differential Scanning; D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D008027:Light; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D055550:Protein Stability; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G
keywords: antibody therapeutics; developability; developable; high-throughput screening; protein aggregation; protein formulation; stability; thermal denaturation