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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Thomas, Douglas D. / Turk, Boris / Wittinghofer, Alfred

IMPACT FACTOR 2017: 3.022

CiteScore 2017: 2.81

SCImago Journal Rank (SJR) 2017: 1.562
Source Normalized Impact per Paper (SNIP) 2017: 0.705

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Volume 393, Issue 8


Protein O-glycosylation analysis

Gerhild Zauner
  • Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, P.O. Box 9600, NL-2300 RC Leiden, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Radoslaw P. Kozak / Richard A. Gardner / Daryl L. Fernandes / André M. Deelder
  • Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, P.O. Box 9600, NL-2300 RC Leiden, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Manfred Wuhrer
  • Corresponding author
  • Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, P.O. Box 9600, NL-2300 RC Leiden, The Netherlands
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar


This review provides an overview on the methods available for analysis of O-glycosylation. Three major themes are addressed: analysis of released O-glycans including different O-glycan liberation, derivatization, and detection methods; analysis of formerly O-glycosylated peptides yielding information on O-glycan attachment sites; analysis of O-glycopeptides, representing by far the most informative but also most challenging approach for O-glycan analysis. Although there are various techniques available for the identification of O-linked oligosaccharides, the focus here is on MS fragmentation techniques such as collision-induced fragmentation, electron capture dissociation, and electron transfer dissociation. Finally, the O-glycan analytical challenges that need to be met will be discussed.

Keywords: electron capture dissociation; electron transfer dissociation; mass spectrometry; O-glycan; O-glycopeptide; O-glycosylation

About the article

Gerhild Zauner

Dr. Gerhild Zauner studied pharmaceutical sciences at the University of Graz (Austria) and the University of East Anglia (UK) and obtained her degree in 2004 before moving to Leiden (The Netherlands) for her PhD in Protein Chemistry/Biophysics at the Chemistry Institute under the supervision of Prof. Gerard W. Canters and Prof. Thijs J. Aartsma. At the beginning of 2009 she started a postdoc at the LUMC in Leiden in the group of Dr. Manfred Wuhrer in the fi eld of glycosylation analysis. Gerhild especially focuses on O-glycosylation analysis, and – together with industrial collaborations – specializes on the development of highthroughput methods for protein glycosylation analysis.

Radoslaw P. Kozak

Radoslaw P. Kozak studied chemistry at Warsaw University, Poland and obtained his Master of Science degree in 2007 working on synthesis of hydrindane precursors of vitamins D with modifi ed side chains. In 2008, he joined Ludger Ltd. where he works as a Scientist and he is involved in chemical glycan release methodology and glycan derivitization by fl uorophore and chromophore tagging for MS and HPLC; glycosylation analysis and glycoprofi ling. In 2010, he started his PhD thesis in collaboration with the Department of Parasitology at Leiden University Medical Centre working on development of analytical methods for O-glycosylation analysis of recombinantly expressed proteins under the supervision of Prof. Andre M. Deelder and Dr. Manfred Wuhrer.

Richard A. Gardner

Dr. Richard A. Gardner studied Biological Chemistry at the University of Leicester before gaining his PhD in Bioorganic Chemistry, also at Leicester. After a postdoctoral position in Organic Chemistry at the University of Central Florida, Orlando, he returned to the UK to work for Evotec OAI in the Development section. After a year developing alternative routes to synthesize final compounds and large scale synthesis of APIs, he moved to his current position as Senior Scientist at Ludger Ltd. The main focus of his activities at Ludger include; research and development into new glycan release and glycan labelling kits, high throughput analysis of glycoproteins and glycans and SPE clean up methodologies.

Daryl L. Fernandes

Dr. Daryl Fernandes is the founder and Chief Executive of Ludger. Daryl has over twenty-nine years experience in development of glycoanalysis methods for glycoproteins of the immune system and biopharmaceuticals. He gained his doctorate at the Glycobiology Institute, University of Oxford and went on to become Process Development Manager and then Head of Analytical Services at Oxford Glycosciences (OGS). Daryl left OGS to found Ludger in 1999. His interests at Ludger include the development and manufacture of reliable biopharmaceutical glycoprofi ling technology and improving the methods for design and production of glycosylated biopharmaceuticals.

André M. Deelder

Prof. Dr. André M. Deelder graduated in Biology in 1971 at the University of Leiden. He performed his PhD research at the department of Parasitology and received his PhD-degree in 1973. In 1985 he was appointed professor in Parasitology and head of the Department of Parasitology at the LUMC. He is leading the Biomolecular Mass Spectrometry Unit with research in clinical proteomics, metabolomics, glycomics and lipidomics. André has published extensively in the fi elds of molecular parasitology, proteomics and applied and translational research.

Manfred Wuhrer

Dr. Manfred Wuhrer studied biochemistry in Regensburg, Germany. He did his PhD (1999) and a subsequent postdoc in the group of Prof. Rudolf Geyer at Giessen University focusing on structural analysis of glycoconjugates. In 2003 he joined the group of Prof. Deelder at Leiden University Medical Center, The Netherlands. He works there currently as an associate professor and is heading the Glycomics and Glycoproteomics group. He is coordinator of the HighGlycan FP7 consortium dealing with the development of highthroughput glycoanalytical techniques for the characterization of biopharmaceuticals as well as for clinical and biological research.

Corresponding author

Received: 2012-06-05

Accepted: 2012-06-05

Published in Print: 2012-08-01

Citation Information: , Volume 393, Issue 8, Pages 687–708, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/hsz-2012-0144.

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