Analysis of glycosaminoglycan‐derived disaccharides in biologic samples by capillary electrophoresis and protocol for sequencing glycosaminoglycans

Abstract

Glycosaminoglycans are biologically significant carbohydrates which either as free chains (hyaluronan) or constituents of proteoglycans (chondroitin/dermatan sulfates, heparin, heparan sulfate and keratan sulfate) participate and regulate several cellular events and (patho)physiological processes. Capillary electrophoresis, due to its high resolving power and sensitivity, has been successfully used for the analysis of glycosaminoglycans. Determination of compositional characteristics, such as disaccharide sulfation pattern, is a useful prerequisite for elucidating the interactions of glycosaminoglycans with matrix effective molecules and, therefore, essential in understanding the biological functions of proteoglycans. The interest in the field of characterization of such biologically important carbohydrates is soaring and advances in this field will signal a new revolution in the area of glycomics equivalent to that of genomics and proteomics. This review focuses on the capillary electrophoresis methods used to determine the disaccharide pattern of glycosaminoglycans in various biologic samples as well as advances in the sequence analysis of glycosaminoglycans using both chromatographic and electrophoretic techniques.

Evolution of glycosaminoglycans and their glycosyltransferases: Implications for the extracellular matrices of animals and the capsules of pathogenic bacteria

“…These reagents readily discern between polymers that differ only in anomeric linkage or epimers. Current methodologies of high-resolution chromatography, mass spectroscopy, fluorophore-assisted electrophoresis, and/or capillary electrophoresis of oligosaccharides generated by specific GAG degradative enzymes have allowed much progress to be made (Imanari et al, 1996;Calabro et al, 2000;Koketsu and Linhardt, 2000;Sturiale et al, 2001;Lamari et al, 2002). However, these analyses of high-molecular-weight polymers still involve fragmentation, separation, and virtual reconstruction to solve the "actual" structure.…”

Chondroitin 6-sulfate as a novel biomarker for mucopolysaccharidosis IVA and VII

“…CS levels have been determined by HPLC, LC-MS/MS, and capillary electrophoresis, based on the differences in enzymatic digestion using chondroitinase ABC and/or chondroitinase ACII (Imanari et al 1996;Koshiishi et al 1998;Oguma et al 2001;Karamanos and Hjerpe 2001;Lamari et al 2002). Although capillary electrophoresis studies provided subjective data that CS is elevated in the urine of MPS IVA (Hopwood and Harrison 1982;Hata and Nagai 1972), C4S and C6S were not separated.…”

Laser‐induced fluorescence as a powerful detection tool for capillary electrophoretic analysis of heparin/heparan sulfate disaccharides

“…The separation principle of the various CE operating modes and the detection methods commonly used in carbohydrate analysis have been extensively reviewed (Mao et al, 2002;Lamari et al, 2002). The disaccharide composition of heparin and HS has been successfully studied with CE using normal or reverse polarity (Ampofo et al, 1991;Damm et al, 1992).…”