High-resolution preparative separation of glycosaminoglycan oligosaccharides by polyacrylamide gel electrophoresis


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Laremore T. N. , Ly M., SOLAKYILDIRIM K. , Zagorevski D. V. , Linhardt R. J.

Analytical Biochemistry, vol.401, no.2, pp.236-241, 2010 (Journal Indexed in SCI Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 401 Issue: 2
  • Publication Date: 2010
  • Doi Number: 10.1016/j.ab.2010.03.004
  • Title of Journal : Analytical Biochemistry
  • Page Numbers: pp.236-241
  • Keywords: ESI-FTMS, Heparin oligosaccharides, Preparative PAGE

Abstract

Separation of milligram amounts of heparin oligosaccharides ranging in degree of polymerization from 4 to 32 is achieved within 6. h using continuous elution polyacrylamide gel electrophoresis (CE-PAGE) on commercially available equipment. The purity and structural integrity of CE-PAGE-separated oligosaccharides are confirmed by strong anion exchange high-pressure liquid chromatography, electrospray ionization Fourier transform mass spectrometry, and two-dimensional nuclear magnetic resonance spectroscopy. The described method is straightforward and time-efficient, affording size-homogeneous oligosaccharides that can be used in sequencing, protein binding, and other structure-function relationship studies. © 2010 Elsevier Inc.