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Boronate affinity saccharide electrophoresis: a novel carbohydrate analysis tool
Authors:Jackson Thomas R  Springall Jeremy S  Rogalle Damien  Masumoto Naoko  Ching Li Hung  D'Hooge François  Perera Semali P  Jenkins A Toby A  James Tony D  Fossey John S  van den Elsen Jean M H
Institution:Department of Biology and Biochemistry, University of Bath, Bath, UK.
Abstract:The incorporation of specialised carbohydrate affinity ligand methacrylamido phenylboronic acid in polyacrylamide gels for fluorophore-assisted carbohydrate electrophoresis greatly improved the effective separation of saccharides that show similar mobilities in standard electrophoresis. Polyacrylamide gel electrophoresis using methacrylamido phenylboronic acid in low loading (typically 0.5-1% dry weight) was unequivocally shown to alter retention of labelled saccharides depending on their boronate affinity. While conventional fluorophore-assisted carbohydrate electrophoresis of 2-aminoacridone labelled glucose oligomers showed an inverted parabolic migration, an undesired trait of small oligosaccharides labelled with this neutral fluorophore, boron affinity saccharide electrophoresis separation of these carbohydrates completely restored their predicted running order, based on their charge/mass ratio, and resulted in improved separation of the analyte saccharides. These results exemplify boron affinity saccharide electrophoresis as an important new technique for analysing carbohydrates and sugar-containing molecules.
Keywords:Boronic acid  Carbohydrate affinity electrophoresis  Fluorophore‐assisted carbohydrate electrophoresis
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