High-throughput profiling of the serum N-glycome on capillary electrophoresis microfluidics systems: toward clinical implementation of GlycoHepatoTest.
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.
Vanderschaeghe
Dieter
D
Unit for Molecular Glycobiology, Department for Molecular Biomedical Research, VIB, Technologiepark 927, B-9052 Ghent, Belgium.
Szekrényes
Akos
A
Wenz
Christian
C
Gassmann
Marcus
M
Naik
Natasha
N
Bynum
Maggie
M
Yin
Hongfeng
H
Delanghe
Joris
J
Guttman
Andras
A
Callewaert
Nico
N
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
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