This content is Free Access.
Chapters
- 00:00Title
- 00:55Introduction
- 01:42Preparing Lectin-Conjugated POROS Columns
- 05:15Packing the Lectin-POROS Beads into a PEEK Column
- 08:11HPLC of Samples
- 11:49Desalting the Collected Fractions and PNGase F-digestion
- 14:50Ziptip Desalting of PNGaseF Samples
- 16:34Glycopeptide Analysis Representative Results
- 19:45Conclusion
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.
Tags
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
