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

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


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 



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).


  1. 1.
    Bravo AA, Sheth SG, Chopra S (2001) Liver biopsy. N Engl J Med 344:495–500PubMedCrossRefGoogle Scholar
  2. 2.
    Colloredo G et al (2003) Impact of liver biopsy size on histological evaluation of chronic viral hepatitis: the smaller the sample, the milder the disease. J Hepatol 39:239–244PubMedCrossRefGoogle Scholar
  3. 3.
    Bedossa P, Dargere D, Paradis V (2003) Sampling variability of liver fibrosis in chronic hepatitis C. Hepatology 38:1449–1457PubMedGoogle Scholar
  4. 4.
    Gressner AM, Gao CF, Gressner OA (2009) Non-invasive biomarkers for monitoring the fibrogenic process in liver: a short survey. World J Gastroenterol 15:2433–2440PubMedCrossRefGoogle Scholar
  5. 5.
    Mas VR et al (2009) Proteomics and liver fibrosis: identifying markers of fibrogenesis. Expert Rev Proteomics 6:421–431PubMedCrossRefGoogle Scholar
  6. 6.
    Callewaert N et al (2004) Noninvasive diagnosis of liver cirrhosis using DNA sequencer-based total serum protein glycomics. Nat Med 10:429–434PubMedCrossRefGoogle Scholar
  7. 7.
    Vanderschaeghe D et al (2009) GlycoFibroTest is a highly performant liver fibrosis biomarker derived from DNA sequencer-based serum protein glycomics. Mol Cell Proteomics 8:986–994PubMedCrossRefGoogle Scholar
  8. 8.
    Gui HL et al (2010) Altered serum N-glycomics in chronic hepatitis B patients. Liver Int 30:259–267PubMedCrossRefGoogle Scholar
  9. 9.
    Chen C et al (2009) Serum protein N-glycans profiling for the discovery of potential biomarkers for nonalcoholic steatohepatitis. J Proteome Res 8:463–470PubMedCrossRefGoogle Scholar
  10. 10.
    Vanderschaeghe D et al (2010) High-throughput profiling of the serum N-glycome on capillary electrophoresis microfluidics systems: toward clinical implementation of GlycoHepatoTest. Anal Chem 82:7408–7415PubMedCrossRefGoogle Scholar
  11. 11.
    Olajos M et al (2008) Sample preparation for the analysis of complex carbohydrates by ­multicapillary gel electrophoresis with light-emitting diode induced fluorescence detection. Anal Chem 80:4241–4246PubMedCrossRefGoogle Scholar
  12. 12.
    Laroy W, Contreras R, Callewaert N (2006) Glycome mapping on DNA sequencing equipment. Nat Protoc 1:397–405PubMedCrossRefGoogle Scholar
  13. 13.

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

Personalised recommendations