Isolation and recovery of acidic oligosaccharides from polyacrylamide gels by semi‐dry electrotransfer

Ali Al‐Hakim, Robert J. Linhardt

Research output: Contribution to journalArticlepeer-review

46 Scopus citations

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.

Original languageEnglish
Pages (from-to)23-28
Number of pages6
JournalElectrophoresis
Volume11
Issue number1
DOIs
StatePublished - 1990
Externally publishedYes

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