Pig mucosal glycosaminoglycans (GAGs) will be systematically fractionated to separate the component heparins, heparan sulfates, and dermatan sulfate according to their anionic functionality by charge density. These component GAGs will also be separated using membrane ultrafiltration into various molecular weight sizes. The GAGs will be extracted from the crude mucosal mixture or as isolated fractions on a charge basis using organic soluble quaternary ammonium salts, salt gradients, and membrane filtration processes. The materials and methods of ultrafiltration and the conditions of the salt/quaternary ammonium-GAGs fractionation will be optimized in Phase I of this project. The fractions will be tested in an animal model for potency in terms of lipoprotein lipase releasing capacity (hypolipidemic activity). Anticoagulant activity and antithrombotic activity will be tested in vitro. Molecular weights with respect to gel permeation chromatography, electrophoretic behavior, chemical composition, component sugars and amino acid content, and sulfate to carboxylate ratios will be assessed for each type of GAGs. Potential use and marketing of these compounds will be as research fine biochemicals to promote new exploration in vascular disease research. Future applications of the novel molecular weight fractions of GAGs will be directed toward assessment of their oral uptake as hypolipidemic or antithrombotic agents and immobilization on replacement organ or vessel materials as biologically active surface modifiers.National Heart, Lung, and Blood Institute (NHLBI)
