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High-Throughput Profiling of the Serum N-Glycome on Capillary Electrophoresis Microfluidics Systems

  • Dieter Vanderschaeghe
  • Andras Guttman
  • Nico CallewaertEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 919)

Abstract

Glycosylation research has gained significant attention in several research fields including immunology, protein production, and biomarker discovery. However, complex and time-consuming protocols are often necessary to obtain suitable samples for analysis. We here describe a short and robust assay to prepare 8-aminopyrene-1,3,6-trisulphonic acid-labeled N-glycans from serum samples. It only involves the subsequent addition of reagents and incubation in a PCR thermocycler. Moreover, this assay allows the detection of these glycans, which are only present in minute amounts in serum, on high-throughput microfluidics CE platforms including the MCE-202 MultiNA, 2100 Bioanalyzer, and eGene system. Using this clinical glycomics assay, we could reliably measure GlycoHepatoTest, a panel of biomarkers allowing the follow-up of chronic liver disease patients from the early stage onward.

Key words

8-Aminopyrene-1,3,6-trisulphonic acid Chronic liver disease GlycoHepatoTest N-glycosylation 

Notes

Acknowledgments

Research was supported by a GOA grant (BOFF07/GOA/017) from Ghent University, by Marie Curie Excellence Grant No. 014292 (EU-Framework Programme 6) and by the Fondation Fournier-Majoie pour l’Innovation (FFMI). Dr. D. Vanderschaeghe is a postdoctoral fellow of FWO (Fonds Wetenschappelijk Onderzoek—Vlaanderen).

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Dieter Vanderschaeghe
    • 1
  • Andras Guttman
    • 2
  • Nico Callewaert
    • 3
    • 4
    • 5
    • 6
    Email author
  1. 1.Department for Molecular Biomedical Research, Unit for Molecular GlycobiologyVIBGhentBelgium
  2. 2.Horvath Laboratory of Bioseparation Sciences, Institute of Medical ChemistryUniversity of DebrecenDebrecenHungary
  3. 3.Department for Molecular Biomedical Research, Unit for Medical BiotechnologyVIB-Ghent UniversityGhent-ZwijnaardeBelgium
  4. 4.Department of Biochemistry and MicrobiologyGhent UniversityGhentBelgium
  5. 5.Department for Molecular Biomedical ResearchUnit for Molecular GlycobiologyGhentBelgium
  6. 6.Department of Biochemistry, Physiology and MicrobiologyGhent UniversityGhentBelgium

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