Large-scale biophysical evaluation of protein PEGylation effects: in vitro properties of 61 protein entities

Research output: Contribution to journalJournal articleResearchpeer-review

Erik Vernet, Gina Popa, Irina Pozdnyakova, Jakob Ewald Rasmussen, Holger Grohganz, Lise Giehm, Malene Hillerup Jensen, Huabing Wang, Bitten Plesner, Hanne Mørck Nielsen, Knud Jørgen Jensen, Jens Berthelsen, Michael Sundström, Marco van de Weert

PEGylation is the most widely used method to chemically modify protein biopharmaceuticals, but surprisingly limited public data is available on the biophysical effects of protein PEGylation. Here we report the first large-scale study, with site-specific mono-PEGylation of 15 different proteins and characterization of 61 entities in total using a common set of analytical methods. Predictions of molecular size were typically accurate in comparison with actual size determined by size-exclusion chromatography (SEC) or dynamic light scattering (DLS). In contrast, there was no universal trend regarding the effect of PEGylation on the thermal stability of a protein based on data generated by circular dichroism (CD), differential scanning calorimetry (DSC), or differential scanning fluorimetry (DSF). In addition, DSF was validated as a fast and inexpensive screening method for thermal unfolding studies of PEGylated proteins. Multivariate data analysis revealed clear trends in biophysical properties upon PEGylation for a subset of proteins, although no universal trends were found. Taken together, these findings are important in the consideration of biophysical methods and evaluation of second-generation biopharmaceutical drug candidates.

Original languageEnglish
JournalMolecular Pharmaceutics
Volume13
Issue number5
Pages (from-to)1587-1598
Number of pages12
ISSN1543-8384
DOIs
Publication statusPublished - 2016

ID: 161942022