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Published on: November 20, 2014
Purification of derivatized oligosaccharides by solid phase extraction for glycomic analysis
Qiwei Zhang1, Henghui Li1, Xiaojun Feng1
1Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics and Molecular Imaging Key Laboratory, Systems Biology Theme, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.
Microcrystalline cellulose SPE cartridges effectively purify derivatized oligosaccharides for sensitive glycan analysis. This method enhances N-glycan profiling in glycoproteins and human serum, showing great potential for glycomics.
Area of Science:
- Analytical Chemistry
- Glycobiology
- Biochemistry
Background:
- Glycan analysis from proteins is complex and crucial for understanding biological processes.
- Chemical derivatization is essential for enhancing carbohydrate detection sensitivity.
- Purification of derivatized oligosaccharides is necessary to remove reagent interference.
Purpose of the Study:
- To compare various solid-phase extraction (SPE) methods for purifying derivatized oligosaccharides.
- To identify the most effective SPE material for clean-up processes in glycan analysis.
- To establish an optimized SPE method for sensitive and accurate N-glycan profiling.
Main Methods:
- Systematic comparison of seven types of self-packed SPE cartridges.
- Optimization of SPE conditions for each cartridge type.
- Analysis of derivatized maltoheptaose using normal-phase HPLC.
- Relative quantification of N-glycans from model glycoproteins and human serum samples.
Main Results:
- Microcrystalline cellulose was identified as the most suitable SPE material for derivatized oligosaccharide purification.
- Optimized SPE method achieved a detection limit of 0.12 pmol for derivatized maltoheptaose with >70% recovery.
- Accurate relative quantification of N-glycans from glycoproteins and determination of over 40 N-glycans in human serum were achieved.
Conclusions:
- Microcrystalline cellulose SPE cartridges offer a simple, reliable, and effective purification strategy for derivatized oligosaccharides.
- The developed method demonstrates high sensitivity and stability, enabling accurate glycan profiling.
- This microcrystalline cellulose-based SPE approach shows significant potential for advancing glycomics research.
