Improving Glycopeptide Fingerprints with High Resolution Chromatography

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Beth L. Gillece-Casto, Thomas E. Wheat, Ziling Lu, and Jeffrey R. Mazzeo [Waters]
Third Symposium on the Practical Applications of Mass Spectrometry in the Biotechnology and Pharmaceutical Industries; Practical MS
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Practical MS
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Characterization of glycoproteins entails analysis of the primary structure of the protein and analysis of the attached oligosaccharides. Oligosaccharides are routinely characterized by glycan mapping after release from the protein. Peptide maps can provide information about the site-specific micro heterogeneity of the glycans if the glycopeptides are resolved.

The enhanced chromatographic resolution associated with Ultra Performance Liquid Chromatography™ has been demonstrated for peptide mapping. The technique improves resolution by a factor of three or more. The Peptide Separation Technology Columns for UPLC™ include both 130Å and 300Å pore size materials. Coupling UPLC™ to oa-Tof mass spectrometry allows high sensitivity identification of glycopeptides through exact mass measurement and deconvolution of complex mass spectra. Optimized chromatographic resolution of individual glycoforms of glycopeptides yields high quality mass spectra. This spectral information can be used to develop UPLC™ methods for quantitative glycopeptide mapping with UV detection.

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