Isolation and Characterization of Heparan Sulfate from Crude Porcine Intestinal Mucosa Peptidoglycan Heparin
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Authors
Griffin, C.C.
Linhardt, Robert J.
VanGorp, C.L.
Toida, T.
Hileman, R.E.
Schubert, R.L.
Brown, S.E.
Issue Date
1995
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Alternative Title
Abstract
A method for the preparation of heparan sulfate from peptidoglycan heparin is described. The objective of this research was to provide a basis for the development and validation of an industrial process to support the preclinical development of heparan sulfate and/or heparan sulfate derivatives. In the preparation of heparan sulfate, heparin was recovered by alcohol fractionation and dermatan sulfate was isolated by selective precipitation. The remaining crude heparan sulfate was fractionated by anion-exchange chromatography into five subfractions. The biological activities of these subfractions were examined by anticoagulant and amidolytic assays. Molecular weight and molecular size were determined using capillary viscometry and polyacrylamide gel electrophoresis. Charge density and degree of sulfation were determined by cellulose acetate electrophoresis and elemental analysis. Oligosaccharide and disaccharide analysis relied on enzymatic depolymerization using heparin lyases followed by polyacrylamide gel and capillary electrophoresis. 1H NMR analysis provided detailed structural information on each subfraction. Crude heparin sulfate and its subfractions showed significant differences in physical, structural and biological properties.
Description
Carbohydrate Research, 276, 183-197
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Full Citation
Isolation and Characterization of Heparan Sulfate from Crude Porcine Intestinal Mucosa Peptidoglycan Heparin, C. C. Griffin, R.J. Linhardt, C.L. VanGorp, T. Toida, R.E. Hileman, R.L. Schubert, S.E. Brown, Carbohydrate Research, 276, 183- 197, 1995.