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Examination of Thermal Unfolding and Aggregation Profiles of a Series of Developable Therapeutic Monoclonal Antibodies
journal contribution
posted on 2015-04-06, 00:00 authored by Mark L. Brader, Tia Estey, Shujun Bai, Roy W. Alston, Karin K. Lucas, Steven Lantz, Pavel Landsman, Kevin M. MaloneyScreening
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.