Boronate affinity saccharide electrophoresis: A novel carbohydrate analysis tool

T R Jackson, J S Springall, D Rogalle, N Masurnoto, H C Li, F D'Hooge, S P Perera, A T A Jenkins, T D James, J S Fossey, J M H van den Elsen

Research output: Contribution to journalArticle

37 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)4185-4191
Number of pages7
JournalElectrophoresis
Volume29
Issue number20
DOIs
Publication statusPublished - 2008

Keywords

  • Carbohydrate affinity electrophoresis
  • Boronic acid
  • Fluorophore-assisted carbohydrate electrophoresis

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