A novel glycosidase plate-based assay for the quantification of galactosylation and sialylation on human IgG

Changes in human IgG galactosylation and sialylation have been associated with several inflammatory diseases which are a major burden on the health care system. A large body of work on well-established glycomic and glycopeptidomic assays has repeatedly demonstrated inflammation-induced changes in Ig...

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Bibliographic Details
Published in:Glycoconjugate journal 2020-12, Vol.37 (6), p.691-702
Main Authors: Rebello, Osmond D., Gardner, Richard A., Urbanowicz, Paulina A., Bolam, David N., Crouch, Lucy I., Falck, David, Spencer, Daniel I. R.
Format: Article
Language:English
Subjects:
Antibodies
Biochemistry
Biomedical and Life Sciences
Chromatography
Dehydrogenases
Galactose
Glycoproteins
Glycosylation
Health care
Immunoglobulin G
Inflammatory bowel disease
Inflammatory diseases
Laboratories
Lectins
Life Sciences
Monosaccharides
Original
Original Article
Pathology
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Summary:Changes in human IgG galactosylation and sialylation have been associated with several inflammatory diseases which are a major burden on the health care system. A large body of work on well-established glycomic and glycopeptidomic assays has repeatedly demonstrated inflammation-induced changes in IgG glycosylation. However, these assays are usually based on specialized analytical instrumentation which could be considered a technical barrier for uptake by some laboratories. Hence there is a growing demand for simple biochemical assays for analyzing these glycosylation changes. We have addressed this need by introducing a novel glycosidase plate-based assay for the absolute quantification of galactosylation and sialylation on IgG. IgG glycoproteins are treated with specific exoglycosidases to release the galactose and/or sialic acid residues. The released galactose monosaccharides are subsequently used in an enzymatic redox reaction that produces a fluorescence signal that is quantitative for the amount of galactosylation and, in-turn, sialylation on IgG. The glycosidase plate-based assay has the potential to be a simple, initial screening assay or an alternative assay to the usage of high-end analytical platforms such as HILIC-FLD-MSn when considering the analysis of galactosylation and sialylation on IgG. We have demonstrated this by comparing our assay to an industrial established HILIC-FLD-MSn glycomic analysis of 15 patient samples and obtained a Pearson’s r correlation coefficient of 0.8208 between the two methods.
ISSN:0282-0080
1573-4986
DOI:10.1007/s10719-020-09953-9