Affordable Access

deepdyve-link
Publisher Website

Building a PGC-LC-MS N-glycan retention library and elution mapping resource.

Authors
  • Abrahams, Jodie L1, 2
  • Campbell, Matthew P1, 2
  • Packer, Nicolle H3, 4
  • 1 Department of Chemistry and Biomolecular Sciences, Faculty of Science & Engineering, Macquarie University, North Ryde, Sydney, NSW, 2109, Australia. , (Australia)
  • 2 Institute for Glycomics, Griffith University, QLD, Gold Coast, 4222, Australia. , (Australia)
  • 3 Department of Chemistry and Biomolecular Sciences, Faculty of Science & Engineering, Macquarie University, North Ryde, Sydney, NSW, 2109, Australia. [email protected] , (Australia)
  • 4 Institute for Glycomics, Griffith University, QLD, Gold Coast, 4222, Australia. [email protected] , (Australia)
Type
Published Article
Journal
Glycoconjugate Journal
Publisher
Springer-Verlag
Publication Date
Sep 13, 2017
Identifiers
DOI: 10.1007/s10719-017-9793-4
PMID: 28905148
Source
Medline
Keywords
License
Unknown

Abstract

Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows.

Report this publication

Statistics

Seen <100 times