title: GlycoFibroTest is a highly performant liver fibrosis biomarker derived from DNA sequencer-based serum protein glycomics.
abstract: Liver fibrosis is currently assessed by liver biopsy, a costly and rather cumbersome procedure that is unsuitable for frequent patient monitoring, which drives research into biomarkers for this purpose. To investigate whether the serum N-glycome contains information suitable for this goal, we developed a 96-well plate-based serum N-glycomics sample preparation protocol that only involves fluid transfer steps and incubations in a PCR thermocycler yielding 8-aminopyrene-1,3,6-trisulfonic acid-labeled N-glycans. These N-glycans are then ready for analysis on the capillary electrophoresis-based DNA sequencers that are the current standard in clinical genetics laboratories worldwide. Subsequently we performed a multicenter, blinded study of 376 consecutive chronic hepatitis C virus patients for which liver biopsies and extensive serum biochemistry data were available. Among patients, the METAVIR fibrosis stage distribution was as follows: 10.6% F0, 44.4% F1, 20.5% F2, 18.4% F3, and 6.1% F4. We found that the ratio of two N-glycans, here called GlycoFibroTest, correlates with the histological fibrosis stage equally well as FibroTest (rho = 0.4-0.5 in F1-F4), which is used in the clinic today. Finally using affinity chromatography we depleted sera of immunoglobulin G, and this resulted in a complete removal of the undergalactosylated biantennary glycans from the N-glycome, which are partially determining GlycoFibroTest.
authors.affiliation: Unit for Molecular Glycobiology, Department for Molecular Biomedical Research, Flanders Institute for Biotechnology (VIB), Technologiepark 927, B-9052 Ghent, Belgium.
authors.forename: Dieter
authors.initials: D
authors.lastname: Vanderschaeghe
authors.forename: Wouter
authors.initials: W
authors.lastname: Laroy
authors.forename: Erwin
authors.initials: E
authors.lastname: Sablon
authors.forename: Philippe
authors.initials: P
authors.lastname: Halfon
authors.forename: Annelies
authors.initials: A
authors.lastname: Van Hecke
authors.forename: Joris
authors.initials: J
authors.lastname: Delanghe
authors.forename: Nico
authors.initials: N
authors.lastname: Callewaert
mesh_terms: D019540:Area Under Curve; D015415:Biomarkers / Q000032:analysis; D001798:Blood Proteins / Q000032:analysis*; D002236:Carbohydrate Conformation; D002908:Chronic Disease; D054794:Glycomics / Q000379:methods*; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G; D008103:Liver Cirrhosis / Q000175:diagnosis* / Q000473:pathology; D011134:Polysaccharides / Q000737:chemistry; D012044:Regression Analysis; D017422:Sequence Analysis, DNA / Q000295:instrumentation*; D000511:alpha-Macroglobulins / Q000032:analysis
chemical_list: D015415:Biomarkers; D001798:Blood Proteins; D007074:Immunoglobulin G; D011134:Polysaccharides; D000511:alpha-Macroglobulins