High-throughput profiling of the serum N-glycome on capillary electrophoresis microfluidics systems

Methods Mol Biol. 2013:919:87-96. doi: 10.1007/978-1-62703-029-8_9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Electrophoresis, Capillary / methods*
  • Glycomics / methods*
  • High-Throughput Screening Assays / methods*
  • Humans
  • Metabolome*
  • Microfluidics / methods*
  • Polysaccharides / blood*
  • Polysaccharides / chemistry
  • Pyrenes / chemistry

Substances

  • 1-aminopyrene-3,6,8-trisulfonic acid
  • Polysaccharides
  • Pyrenes