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Using micro pillar array columns (μPAC) for the analysis of permethylated glycans
Byeong Gwan Cho1, Peilin Jiang1, Mona Goli1
1Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409-1061, USA. yehia.mechref@ttu.edu.
The Analyst
|June 16, 2021
Summary
Micro array pillar nanoLC columns (μPAC) show promise for analyzing permethylated glycans. These columns offer efficient separation and purification, advancing glycan analysis in biological systems.
Area of Science:
- Biochemistry
- Analytical Chemistry
Background:
- Glycosylation is a crucial post-translational protein modification.
- Liquid chromatography-mass spectrometry (LC-MS) is vital for glycan analysis.
- Permethylated glycans are commonly separated using reverse-phase columns.
Purpose of the Study:
- To evaluate the suitability of micro array pillar nanoLC columns (μPAC) for permethylated glycan analysis.
- To compare μPAC performance against established methods and columns.
Main Methods:
- Utilized μPAC trapping columns for online purification, comparing them to PepMap columns.
- Analyzed permethylated N- and O-glycans using 50 cm μPAC columns.
- Investigated the separation capabilities of 200 cm μPAC columns for complex glycans.
Main Results:
- μPAC columns demonstrated effective online purification capabilities.
- 50 cm μPAC columns successfully analyzed both N- and O-glycans.
- 200 cm μPAC columns provided efficient separation of glycan isomers, including oligomannose.
Conclusions:
- μPAC columns are a viable alternative for permethylated glycan analysis.
- The low back pressure, reproducibility, and durability of μPAC columns are advantageous.
- μPAC technology enhances the capabilities of LC-MS for detailed glycan structural characterization.
