Endoglycosidase S Enables a Highly Simplified Clinical Chemistry Procedure for Direct Assessment of Serum IgG Undergalactosylation in Chronic Inflammatory Disease.

Autor:	Vanderschaeghe D; VIB Center for Medical Biotechnology, Technologiepark 927, B-9052 Ghent, Belgium; Department of Biochemistry and Microbiology, Ghent University, Ledeganckstraat 35, B-9000 Ghent, Belgium., Meuris L; VIB Center for Medical Biotechnology, Technologiepark 927, B-9052 Ghent, Belgium; Department of Biochemistry and Microbiology, Ghent University, Ledeganckstraat 35, B-9000 Ghent, Belgium., Raes T; VIB Center for Medical Biotechnology, Technologiepark 927, B-9052 Ghent, Belgium; Department of Biochemistry and Microbiology, Ghent University, Ledeganckstraat 35, B-9000 Ghent, Belgium., Grootaert H; VIB Center for Medical Biotechnology, Technologiepark 927, B-9052 Ghent, Belgium; Department of Biochemistry and Microbiology, Ghent University, Ledeganckstraat 35, B-9000 Ghent, Belgium., Van Hecke A; VIB Center for Medical Biotechnology, Technologiepark 927, B-9052 Ghent, Belgium; Department of Biochemistry and Microbiology, Ghent University, Ledeganckstraat 35, B-9000 Ghent, Belgium., Verhelst X; Laboratory of Hepatology, Department of Hepatology and Gastroenterology, Ghent University Hospital, Corneel Heymanslaan 10, B-9000 Ghent, Belgium., Van de Velde F; Department of Endocrinology, Ghent University Hospital, Corneel Heymanslaan 10, B-9000 Ghent, Belgium., Lapauw B; Department of Endocrinology, Ghent University Hospital, Corneel Heymanslaan 10, B-9000 Ghent, Belgium., Van Vlierberghe H; Laboratory of Hepatology, Department of Hepatology and Gastroenterology, Ghent University Hospital, Corneel Heymanslaan 10, B-9000 Ghent, Belgium., Callewaert N; VIB Center for Medical Biotechnology, Technologiepark 927, B-9052 Ghent, Belgium; Department of Biochemistry and Microbiology, Ghent University, Ledeganckstraat 35, B-9000 Ghent, Belgium. Electronic address: Nico.Callewaert@ugent.vib.be.
Jazyk:	angličtina
Zdroj:	Molecular & cellular proteomics : MCP [Mol Cell Proteomics] 2018 Dec; Vol. 17 (12), pp. 2508-2517. Date of Electronic Publication: 2018 Sep 06.
DOI:	10.1074/mcp.TIR118.000740
Abstrakt:	Over the past 30 years, it has been firmly established that a wide spectrum of (autoimmune) diseases such as rheumatoid arthritis, Crohn's and lupus, but also other pathologies like alcoholic and non-alcoholic steatohepatitis (ASH and NASH) are driven by chronic inflammation and are hallmarked by a reduced level of serum IgG galactosylation. IgG (under)galactosylation is a promising biomarker to assess disease severity, and monitor and adjust therapy. However, this biomarker has not been implemented in routine clinical chemistry because of a complex analytical procedure that necessitates IgG purification, which is difficult to perform and validate at high throughput. We addressed this issue by using endo-β-N-acetylglucosaminidase from Streptococcus pyogenes (endoS) to specifically release Fc N-glycans in whole serum. The entire assay can be completed in a few hours and only entails adding endoS and labeling the glycans with APTS. Glycans are then readily analyzed through capillary electrophoresis. We demonstrate in two independent patient cohorts that IgG undergalactosylation levels obtained with this assay correlate very well with scores calculated from PNGaseF-released glycans of purified antibodies. Our new assay allows to directly and specifically measure the degree of IgG galactosylation in serum through a fast and completely liquid phase protocol, without the requirement for antibody purification. This should help advancing this biomarker toward clinical implementation. (© 2018 Vanderschaeghe et al.)
Databáze:	MEDLINE
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