Isolation and characterization of β-cyclodextrin sulfates by preparative gradient polyacrylamide gel electrophoresis, capillary electrophoresis and electrospray ionization - Mass spectrometry

Ronald E. Hileman, Marshall M. Siegel, Keiko Tabei, Kuberan Balagurunathan, Robert J. Linhardt

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

A β-cyclodextrin sulfate mixture has been fractionated using discontinuous gradient polyacrylamide gel electrophoresis. Semidry electrotransfer of the sample onto a positively charged nylon membrane and visualization of a portion of this membrane with Alcian blue stain showed multiple bands. The bands were cut from the remaining portion of the membrane and after washing with 8 M urea, the β-cyclodextrin sulfate fractions were eluted with 2 M sodium chloride and dialyzed. Analysis of each fraction using high resolution analytical gradient polyacrylamide gel electrophoresis as well as capillary electrophoresis, using indirect detection, showed some of the fractions to be pure while others were mixtures. Each β-cyclodextrin sulfate fraction was complexed with a basic synthetic peptide and analyzed by electrospray ionization mass spectrometry to define the mass of the components in each mixture and thereby to determine the purity of each sample.

Original languageEnglish
Pages (from-to)2677-2681
Number of pages5
JournalElectrophoresis
Volume19
Issue number15
DOIs
StatePublished - Nov 1998
Externally publishedYes

Keywords

  • Capillary electrophoresis
  • Cyclodextrin sulfate
  • Electrospray ionization
  • Glycosaminoglycan
  • Mass spectrometry
  • Polyacrylamide gel electrophoresis

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