dermatan sulfate from beef mucosa - structure, physicochemical and biological properties of fractions prepared by chemical depolymerization and anion-exchange chromatography

Dermatan sulfate was extracted and purified from beef intestinal mucosa. The structure and physicochemical properties were evaluated by different techniques, such as, disaccharide pattern, relative molecular mass, sulfate-to-carboxyl ratio, and electrophoretic profile in agarose electrophoresis. The biological activity was evaluated as heparin cofactor II activity (HCII activity). The purity of dermatan sulfate was carefully evaluated by specific enzymatic cleavage, agarose electrophoresis, and HPLC. Different relative molecular masses of dermatan sulfate, from 25 000 to 2000, were prepared by chemical degradation. The structures and physicochemical properties were checked to exclude a possible desulfation process. The HCII activities were evaluated for different relative molecular mass of dermatan sulfate. The capacity of chondroitinase ABC to cleave different relative molecular masses of dermatan sulfate was also studied. Native dermatan sulfate was fractionated according to charge density. Different fractions were obtained and analysed for disaccharide pattern, relative molecular mass, sulfate-to-carboxyl ratio, and HCII activities.