Effect of Pegylation on Self-Association of IFN-α2b
Angela Mohs
Alexandre Ambrogelly
Xiaoyu Yang
Mark Haverick
Jason K. Cheung
Chakravarthy Narasimhan
Mohammed Shameem
10.1021/mp400343b.s001
https://acs.figshare.com/articles/journal_contribution/Effect_of_Pegylation_on_Self_Association_of_IFN_2b/2335936
Pegylation
of therapeutic proteins is an established technology used to enhance
the bioavailability of an active pharmaceutical ingredient in the
body of patients. While the physiochemical properties of pegylated
monomeric proteins have been extensively described, there is still
limited information on the characterization of pegylated oligomeric
proteins. In this study, we report the characterization of a pegylated
interferon alpha2b (PEGIFN-α2b) concentration-dependent oligomerization
by a series of orthogonal biochemical and biophysical methods. These
methods include sedimentation velocity and sedimentation equilibrium
analytical ultracentrifugation, matrix-assisted laser desorption ionization,
and size exclusion chromatography of bissulfosuccinimidyl suberate
cross-linked PEGIFN. We report here that PEGIFN-α2b self-associates
in a concentration-dependent manner into mainly monomers, dimers,
and trimers. In the presence of the chemical cross-linker, PEGIFN-α2b
is primarily monomeric (57%) at concentration lower than 0.3 mg/mL
and contains about equal amount of monomers and dimers (47.0% and
37.7%, respectively), about 15% of trimers, and up to 4% of higher
molecular weight species at 0.7 mg/mL and above.
2014-01-06 00:00:00
size exclusion chromatography
characterization
pegylated interferon
pegylated oligomeric proteins
dimers
IFN
PEGIFN
weight species
method
sedimentation velocity
trimer
mg
pegylated monomeric proteins
sedimentation equilibrium