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Chapters
- 00:05Title
- 01:10Protein and Glycoprotein Denaturation and Alkylation
- 02:15N-linked Glycan Enzymatic Digestion
- 03:29Elution of N-glycans and Protein Filter Aided Sample Preparation (FASP) Digestion
- 06:37Derivatization of N-linked Glycans with Hydrophobic Hydrazide Tags
- 08:19Ultra-high Pressure Liquid Chromatography and Mass Spectrometry Analysis
- 09:27Results: Analysis of the Glycomics and Proteomics Combined Purification Strategy
- 10:44Conclusion
A high-throughput protocol was developed for combined proteomics and glycomics purification and LC-MS/MS quantification in plasma. Deamidation analysis of N-linked glycosylation motifs was specific to deglycosylated sites. Accurate quantitation of N-glycans was achieved by coupling filter aided N-glycan separation to the individuality normalization when labeling with glycan hydrazide tags strategy.
Tags
Quantitative GlycomicsProteomicsPurification StrategyN-glycansProteinsMass Spectrometry AnalysisScalabilitySpeedAnalytical ReproducibilityGlycomicsGlycosylation Site OccupancyHigh-throughput Biological StudiesN-glycan MeasurementsMass Spectrometry Tailored Derivatization StrategyRelative QuantificationCancer And Control StatesAnalytical VariabilityBiological InteractionsGlycomeProteomeDisease StatesBiomarker Discovery
