Strategic Implementation of Glycan Standards for Reliable Analysis of Glycomolecules

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1st EuroCarb Young Researcher (ECYR) Workshop

CarboMet Workshop

Mini symposium: Glycobiology at the LUMC and beyond

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Strategic Implementation of Glycan Standards for Reliable Analysis of Glycomolecules

Jenifer L. Hendel 1 , Paulina A. Urbanowicz 1 , Radoslaw P. Kozak 1

1 Ludger Ltd., Oxford, United Kingdom

For most therapeutic glycoproteins, glycosylation can greatly influence clinical performance of the drug product, particularly its in vivo safety and efficacy profile.¹ In biological tissues, changes in glycosylation have been associated with the state of health and/or disease of the individual.² Given this, there is increasing interest in accurately characterising glycosylation, for example monitoring glycosylation patterns of biopharmaceutical therapeutics throughout the product life-cycle as well as in glycan biomarker discovery for medical diagnostics.

Robust analytical strategies are required to meet the challenge of accurately and reliably characterising glycosylation. A key component in a well-designed analytical strategy is the inclusion of glycan standards. This can improve glycan characterisation, allow quantitation, and provide vital information on analytical performance. These standards include reference standards, quantitative standards, and process and system suitability standards, respectively. This poster focuses on the uses of well-characterised glycan, glycopeptide and glycoprotein standards to support analysis of glycosylation patterns. We will demonstrate which standards can be used for best practice during the analysis of sialic acids, monosaccharides, N-glycans and O-glycans.

Finally, we will highlight the use of specific glycan and glycoprotein standards throughout a detailed glycan characterisation workflow. As a case study, we will describe the strategy and criteria for using system suitability standards, process standards and reference standards together to ensure reliable analytical data is obtained during endoglycosidase (PNGaseF) release, fluorophore labelling, exoglycosidase sequencing, sample clean-up and UPLC analysis.

P. Zhang, S. Woen, T. Wang, B. Liau, S. Zhao, C. Chen, Y. Yang, Z. Song, M.R. Wormald, C. Yu, P.M. Rudd. Drug Discov Today, 2016,21,740-765.

a) G. Lauc, M. Pezer, I. Rudan, H. Campbell. Biochim Biophys Acta - Gen Subj, 2016,1860,1574-1582. b) B. Adamczyk, T. Tharmalingam, P.M. Rudd. Biochim Biophys Acta - Gen Subj, 2012,1820,1347-1353. c) A. Varki. Glycobiology, 2017,27,3-49.

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