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A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples
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October 01, 2009
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Lectin-conjugated POROS beads were employed for HPLC. Glycopeptide standards served as positive and negative controls. MARS-14 depleted, trypsin-digested human plasma was chromatographed and flow-through (FT) and bound fractions collected for ESI-LC-MS/MS analyses. Glycopeptides were enriched in the bound fraction as compared to FT.
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Lectin HPLC MethodSelectively-glycosylated PeptidesComplex Biological SamplesGlycansPost-translational ModificationsTumor CellsCancer-associated Glycan VariantsBiomarker DevelopmentLectinsSambucus Nigra (SNA)Aleuria Aurantia (AAL)Affinity MatricesPOROS BeadsPEEK ColumnsHigh Pressure Liquid Chromatography (HPLC)Plasma DepletionMultiple Affinity Removal System (MARS-14)Trypsin-digestionFlow-through FractionBound FractionPNGaseFN-linked GlycansLC-MS/MS AnalysisQStar EliteMascot SoftwarePositive ControlsNegative Controls
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Johansen, E., Schilling, B., Lerch, M., Niles, R. K., Liu, H., Li, B., Allen, S., Hall, S. C., Witkowska, H. E., Regnier, F. E., Gibson, B. W., Fisher, S. J., Drake, P. M. A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples. J. Vis. Exp. (32), e1398, doi:10.3791/1398 (2009).
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