O-Linked Oligosaccharide Profiling by HPLC
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The several different core regions of O-linked chains (Chapter 105) can be further extended by Gal and GlcNAc containing backbones or by addition of blood group antigen-type glycosylation and the presence of sialic acid or sulfate. We sought a universal column that can be applied with high resolution to the various oligosaccharide alditols released from glycoproteins by β-elimination (see Note 1) and have pioneered (1,2) the use of porous graphitized carbon (PGC). This is an alternative to C18 reversed-phase HPLC and normal-phase amino-bonded columns, which can be used together in the presence and absence of high-salt buffers (3).
KeywordsSialic Acid Control Software Porous Graphitized Carbon Longe Retention Time Dynamic Mixer
- 2.Davies, M. J., Smith, K. D., Carruthers, R. A., Chai, W., Lawson, A. M., and Hounsell, E. F. (1993) The use of a porous graphitized carbon (PGC) column for the HPLC of oligosaccharides, alditols and glycopeptides with subsequent mass spectrometry analysis. J. Chromatogr. 646, 317–326.PubMedCrossRefGoogle Scholar
- 4.Hounsel, E. F., ed. (1993) Methods in Molecular Biology, vol 14: Glycoprotein Analysis in Biomedicine, Humana, Totowa, NJ.Google Scholar