High-performance gel permeation chromatography of glycosaminoglycans. Column calibration by gel electrophoresis.
High-performance gel permeation chromatography and sensitivity-enhanced polyacrylamide gel electrophoresis (SE-PAGE) have been combined to investigate the chromatographic properties of structurally related glycosaminoglycans. Enzymatically digested samples of hyaluronate, chondroitin 4-sulfate and dermatan sulfate were fractionated on a column series of TSK G4000SW and TSK G2000SW, eluted with 0.15 M sodium chloride. Isolated fractions were subjected to sensitivity-enhanced polyacrylamide gel electrophoresis for determination of molecular weight. These procedures allowed the rapid development of column calibration profiles for each glycosaminoglycan, under a given set of chromatographic conditions. The profiles obtained in 0.15 M sodium chloride yielded the following data: (1) at equal degrees of polymerization, hyaluronic acid chains have an apparently smaller hydrodynamic radius than the sulfated polymers, and (2) at equal molecular weights, all three glycosaminoglycans have approximately equal hydrodynamic radii.
Hittner
D M
DM
Department of Chemistry, Polytechnic University, Brooklyn, NY 11201.
Cowman
M K
MK
D002850:Chromatography, Gel; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D006025:Glycosaminoglycans / Q000032:analysis*; D008970:Molecular Weight
D006025:Glycosaminoglycans