High-resolution preparative separation of glycosaminoglycan oligosaccharides by polyacrylamide gel electrophoresis.
Separation of milligram amounts of heparin oligosaccharides ranging in degree of polymerization from 4 to 32 is achieved within 6h 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.
Laremore
Tatiana N
TN
Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
Ly
Mellisa
M
Solakyildirim
Kemal
K
Zagorevski
Dmitri V
DV
Linhardt
Robert J
RJ
D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel / Q000191:economics / Q000379:methods*; D006493:Heparin / Q000302:isolation & purification*; D019906:Nuclear Magnetic Resonance, Biomolecular; D009844:Oligosaccharides / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization
D009844:Oligosaccharides; D006493:Heparin