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Mol Cell Proteomics. 2014 Jun;13(6):1598-610. doi: 10.1074/mcp.M113.037465. Epub 2014 Apr 9.

Comparative performance of four methods for high-throughput glycosylation analysis of immunoglobulin G in genetic and epidemiological research.

Author information

  • 1From the ‡MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK;
  • 2§Genos Glycoscience Laboratory, Zagreb, Croatia;
  • 3¶Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany; ‖glyXera GmbH, Magdeburg, Germany;
  • 4**Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands;
  • 5‖glyXera GmbH, Magdeburg, Germany;
  • 6‡‡Faculty of Medicine, University of Split, Split, Croatia;
  • 7§§University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, Croatia;
  • 8¶¶Centre for Population Health Sciences, School of Medicine and Veterinary Medicine, University of Edinburgh, Edinburgh, UK;
  • 9¶Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany; ‖‖Otto-von-Guericke University, Chair of Bioprocess Engineering, Magdeburg, Germany;
  • 10Institute of Cytology and Genetics SB RAS, Novosibirsk, Russia; "Yurii Aulchenko" consulting, Groningen, The Netherlands;
  • 11**Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands; Division of BioAnalytical Chemistry, VU University Amsterdam, Amsterdam, The Netherlands.
  • 12§Genos Glycoscience Laboratory, Zagreb, Croatia; §§University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, Croatia; glauc@pharma.hr.

Abstract

The biological and clinical relevance of glycosylation is becoming increasingly recognized, leading to a growing interest in large-scale clinical and population-based studies. In the past few years, several methods for high-throughput analysis of glycans have been developed, but thorough validation and standardization of these methods is required before significant resources are invested in large-scale studies. In this study, we compared liquid chromatography, capillary gel electrophoresis, and two MS methods for quantitative profiling of N-glycosylation of IgG in the same data set of 1201 individuals. To evaluate the accuracy of the four methods we then performed analysis of association with genetic polymorphisms and age. Chromatographic methods with either fluorescent or MS-detection yielded slightly stronger associations than MS-only and multiplexed capillary gel electrophoresis, but at the expense of lower levels of throughput. Advantages and disadvantages of each method were identified, which should inform the selection of the most appropriate method in future studies.

PMID:
24719452
PMCID:
PMC4047478
DOI:
10.1074/mcp.M113.037465
[PubMed - indexed for MEDLINE]
Free PMC Article
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