title: High-throughput profiling of the serum N-glycome on capillary electrophoresis microfluidics systems: toward clinical implementation of GlycoHepatoTest.
abstract: We developed a 3 h procedure for preparing serum N-glycans and labeling them with 8-aminopyrene-1,3,6-trisulfonic acid (APTS) by sequential addition of reagents to the serum and incubation in a polymerase chain reaction (PCR) thermocycler. Moreover, we succeeded in analyzing these samples by capillary electrophoresis on three commercial microfluidics-based platforms: the MCE-202 MultiNA, the 2100 Bioanalyzer, and a modified prototype of the eGene system which were originally designed for nucleic acid separation and detection. Although these instruments use short separation channels, our technical improvements made it possible to reliably measure the N-glycans constituting GlycoHepatoTest. This test comprises a panel of biomarkers that allows follow-up of liver fibrosis patients starting from the early stage. In this way and for the first time, we demonstrate a clinical glycomics assay on an affordable, robust platform so that clinical chemistry laboratories can exploit glycomics in the diagnosis and monitoring of chronic liver disease. Another potential application is the rapid screening of the N-glycosylation of recombinant glycoproteins produced for pharmaceutical use.
authors.affiliation: Unit for Molecular Glycobiology, Department for Molecular Biomedical Research, VIB, Technologiepark 927, B-9052 Ghent, Belgium.
authors.forename: Dieter
authors.initials: D
authors.lastname: Vanderschaeghe
authors.forename: Akos
authors.initials: A
authors.lastname: Szekrényes
authors.forename: Christian
authors.initials: C
authors.lastname: Wenz
authors.forename: Marcus
authors.initials: M
authors.lastname: Gassmann
authors.forename: Natasha
authors.initials: N
authors.lastname: Naik
authors.forename: Maggie
authors.initials: M
authors.lastname: Bynum
authors.forename: Hongfeng
authors.initials: H
authors.lastname: Yin
authors.forename: Joris
authors.initials: J
authors.lastname: Delanghe
authors.forename: Andras
authors.initials: A
authors.lastname: Guttman
authors.forename: Nico
authors.initials: N
authors.lastname: Callewaert
mesh_terms: D015415:Biomarkers / Q000097:blood; D019075:Electrophoresis, Capillary / Q000379:methods*; D054794:Glycomics; D006801:Humans; D008103:Liver Cirrhosis / Q000175:diagnosis / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D009439:Neuraminidase / Q000378:metabolism; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D011134:Polysaccharides / Q000097:blood*; D011721:Pyrenes / Q000737:chemistry
chemical_list: C103430:8-aminopyrene-1,3,6-trisulfonic acid; D015415:Biomarkers; D011134:Polysaccharides; D011721:Pyrenes; D009439:Neuraminidase; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase