Isolation and recovery of acidic oligosaccharides from polyacrylamide gels by semi-dry electrotransfer. |
| |
Authors: | A al-Hakim R J Linhardt |
| |
Institution: | Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242. |
| |
Abstract: | Acidic oligosaccharides derived from glycosaminoglycan heparin were separated by polyacrylamide gradient gel electrophoresis (PAGE). The gel could be visualized using Alcian Blue dye to give a pattern of highly resolved, well defined bands. The particular banding pattern obtained was the result of a heparinase catalyzed depolymerization which afforded oligosaccharide products that differed in size by one disaccharide unit. The separated oligosaccharides could be recovered prior to staining by electroelution onto a positively charged nylon membrane by a semi-dry transfer procedure. Subsequent elution and quantitative recovery of individual oligosaccharides from the membrane was achieved. By using multiple membrane layers a second separation dimension was obtained, resulting in increased oligosaccharide purity proportional to transfer depth. Preparative gradient polyacrylamide gel electrophoresis followed by semi-dry electro-transfer and recovery represents a novel method for the preparation of homogeneous acidic oligosaccharides. |
| |
Keywords: | |
|
|