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Human serum processing and analysis methods for rapid and reproducible N-glycan mass profiling
Scott R Kronewitter1, Maria Lorna A de Leoz, Kyle S Peacock
1Department of Chemistry, University of California Davis, Davis, California 95616, USA.
Journal of Proteome Research
|August 12, 2010
Summary
This study enhances glycan biomarker discovery by improving sample preparation repeatability. We explored microwave-assisted enzymatic release and automated extraction to minimize variability for robust mass spectrometry analysis.
Area of Science:
- Biochemistry and Molecular Biology
- Analytical Chemistry
- Biomarker Discovery
Background:
- Glycans are crucial in biological processes and disease, making them promising biomarkers.
- Glycosylation changes are linked to cancer development, necessitating reliable glycan analysis.
- Current glycan analysis methods require optimization for biomarker discovery due to process variability.
Purpose of the Study:
- To evaluate methods for expediting glycan release and enrichment.
- To identify and minimize variability in glycan sample preparation for robust biomarker discovery.
- To assess the suitability of microwave reactors and automated extraction for glycan analysis.
Main Methods:
- Enzymatic release of glycans using a microwave reactor.
- Automated solid-phase extraction of glycans with a robotic liquid handler.
- Analysis of glycans using matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS).
Main Results:
- Microwave-assisted enzymatic release and automated extraction show potential for reducing sample preparation time and variability.
- System-wide data analysis methods are explored to establish performance metrics for glycan sample preparation.
- Understanding measurement device variability is key to improving overall process robustness.
Conclusions:
- Optimized glycan sample preparation is essential for reliable biomarker discovery.
- Expedited methods like microwave reactors and automation can enhance throughput and reproducibility.
- Further development of data analysis metrics is needed to validate glycan preparation platforms.
