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Molecular & Cellular Proteomics 8:986-994, 2009.
© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

GlycoFibroTest Is a Highly Performant Liver Fibrosis Biomarker Derived from DNA Sequencer-based Serum Protein Glycomics^*,S

Dieter Vanderschaeghe^,, Wouter Laroy^,¶,||, Erwin Sablon^**, Philippe Halfon, Annelies Van Hecke^,¶, Joris Delanghe and Nico Callewaert^,,¶¶

From the Unit for Molecular Glycobiology, Department for Molecular Biomedical Research, Flanders Institute for Biotechnology (VIB), Technologiepark 927, B-9052 Ghent, Belgium, Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, De Pintelaan 185, B-9000 Ghent, Belgium, ^¶ Department of Molecular Biology, Ghent University, Technologiepark 927, B-9052 Ghent, Belgium, ^** Infectious Diseases Unit, Diagnostics R&D, Innogenetics NV, Industriepark Zwijnaarde 7, Box 4, B-9052 Ghent, Belgium, Department of Virology, Alphabio Laboratory, 23 rue de Friedland, 13006 Marseille, France, and Department of Biochemistry, Physiology, and Microbiology, Ghent University, K. L.-Ledeganckstraat 35, B-9000 Ghent, Belgium

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 ( = 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.

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