UNIGE document Scientific Article
previous document  unige:115743  next document
add to browser collection
Title

Orthogonal Middle-up Approaches for Characterization of the Glycan Heterogeneity of Etanercept by Hydrophilic Interaction Chromatography Coupled to High-Resolution Mass Spectrometry

Authors
Nováková, Lucie
Stoll, Dwight
Lauber, Matthew
Beck, Alain
Published in Analytical Chemistry. 2019, vol. 91, no. 1, p. 873-880
Abstract Etanercept is a recombinant Fc fusion protein widely used to treat rheumatic diseases. This protein is highly glycosylated and contains numerous O- and N-glycosylation sites. Since glycosylation is recognized as an important critical quality attribute (CQA) that can affect immunogenicity, solubility, and stability of Fc fusion proteins, it should be thoroughly characterized. In this work, hydrophilic interaction chromatography (HILIC) was combined with high-resolution mass spectrometry (HRMS) by using a quadrupole time-of flight mass spectrometer to assess glycosylation of etanercept at the middle-up level of analysis (fragments of ca. 25−30 kDa). In addition, a combination of different enzymatic digestion procedures (i.e., glycosidase, sialidase, and protease) was systematically employed to facilitate spectra deconvolution. With the developed procedure, the main post-translational modifications (PTMs) of etanercept were assessed, and a global overview of the subunit-specific distribution of the glycosylation pattern was obtained at a middle-up level of analysis.
Identifiers
PMID: 30512936
Full text
Structures
Research group Sciences analytiques
Citation
(ISO format)
D'ATRI, Valentina et al. Orthogonal Middle-up Approaches for Characterization of the Glycan Heterogeneity of Etanercept by Hydrophilic Interaction Chromatography Coupled to High-Resolution Mass Spectrometry. In: Analytical Chemistry, 2019, vol. 91, n° 1, p. 873-880. https://archive-ouverte.unige.ch/unige:115743

111 hits

0 download

Update

Deposited on : 2019-04-04

Export document
Format :
Citation style :