Glycosaminoglycan Chain Analysis and Characterization (Glycosylation/Epimerization)

Glycosaminoglycans (GAGs) including chondroitin sulfate (CS), dermatan sulfate (DS), and heparan sulfate/heparin (HS/Hep) are linear polysaccharides and involved in the regulation of various biological events through interaction with functional proteins. GAGs are modified by sulfation at various positions of each saccharide residue and the epimerization of uronic acid residues during the chain’s biosynthesis, resulting in enormous structural diversity. This structural diversity is the basis for the wide range of biological activities of GAGs. Thus, the structural analysis of GAGs is key to understanding their biological functions. This chapter describes detailed instructions for the extraction and structural analysis of GAGs from cultured cells and tissues using a combination of GAG-degrading enzymes and high-performance liquid chromatography.