Determination of the degree of PEGylation of protein bioconjugates using data from proton nuclear magnetic resonance spectroscopy

Ahlem Zaghmi; Andrea A Greschner; Eduardo Mendez-Villuendas; Jun Yang Liu; Hendrick W de Haan; Marc A Gauthier

doi:10.1016/j.dib.2019.104037

Determination of the degree of PEGylation of protein bioconjugates using data from proton nuclear magnetic resonance spectroscopy

Data Brief. 2019 May 23:25:104037. doi: 10.1016/j.dib.2019.104037. eCollection 2019 Aug.

Authors

Ahlem Zaghmi¹, Andrea A Greschner¹, Eduardo Mendez-Villuendas², Jun Yang Liu¹, Hendrick W de Haan², Marc A Gauthier¹

Affiliations

¹ Institut National de la Recherche Scientifique (INRS), EMT Research Center, Varennes, QC, J3X 1S2, Canada.
² University of Ontario Institute of Technology, Faculty of Science, Oshawa, Ontario, L1H 7K4, Canada.

Abstract

The average number of methoxy poly(ethylene glycol) (mPEG) chains grafted to a protein - also known as the degree of PEGylation - is a fundamental parameter for characterizing a bioconjugate. The degree of PEGylation is typically determined by chromatographic or electrophoretic methods, which are subject to certain biases. This contribution describes an analytical approach alongside technical precautions for quantitatively determining the degree of PEGylation of protein bioconjugates by ¹H NMR spectroscopy. An accompanying dataset, corresponding to the raw ¹H NMR spectra of thirteen bioconjugates with different degrees of PEGylation and different mPEG molecular weights, is provided for the reader to become familiar with the analysis. The exemplary bioconjugate system used in this Data article is the enzyme glutamate dehydrogenase (GDH) modified with multiple copies of mPEG (0.5-20 kDa). These bioconjugates correspond to those discussed in-depth in the article "Mechanisms of activity loss for a multi-PEGylated protein by experiment and simulation" by Zaghmi et al., 2019 The described approach to calculate degree of PEGylation is quantitative, applicable to other proteins, and can be adapted to other types of polymers.