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Electrospray Ionization (ESI) Mass Spectrometry01:12

Electrospray Ionization (ESI) Mass Spectrometry

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ESI utilizes electrical energy to transfer ions from the liquid phase of the sample into the...

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A Quantitative Glycomics and Proteomics Combined Purification Strategy
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Published on: March 8, 2016

Improving N-glycan coverage using HPLC-MS with electrospray ionization at subambient pressure.

Ioan Marginean1, Scott R Kronewitter, Ronald J Moore

  • 1Biological Sciences Division, Pacific Northwest National Laboratory, P.O. Box 999, Richland, Washington 99352, United States.

Analytical Chemistry
|October 3, 2012
PubMed
Summary

Subambient pressure ionization with nanoelectrospray mass spectrometry (SPIN-MS) enhances human serum glycan profiling by improving ionization efficiency and detecting previously unobserved glycans, aiding biomarker discovery.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biomarker Discovery

Background:

  • Human serum glycan profiling using mass spectrometry (MS) shows promise for clinical biomarker discovery.
  • Challenges in comprehensive profiling include low glycan ionization efficiency, large dynamic concentration ranges, and problematic fragmentation of large glycans.

Purpose of the Study:

  • To investigate the utility of subambient pressure ionization with nanoelectrospray mass spectrometry (SPIN-MS) for expanding human serum glycome profiling.
  • To compare SPIN-MS with conventional atmospheric pressure electrospray ionization mass spectrometry (ESI-MS) for enhanced glycan detection.

Main Methods:

  • Liquid chromatographic separation on graphite columns was performed.
  • Subambient pressure ionization with nanoelectrospray mass spectrometry (SPIN-MS) was utilized.
  • Comparison was made with atmospheric pressure electrospray ionization mass spectrometry (ESI-MS) using a heated capillary inlet.

Main Results:

  • SPIN-MS expanded the serum glycome profile compared to conventional ESI-MS.
  • Higher charge states were observed for approximately half of the annotated glycans using SPIN-MS.
  • 34 out of 130 detected glycans were exclusively identified using SPIN-MS, improving coverage of glycan families and larger glycans.

Conclusions:

  • SPIN-MS significantly improves the coverage and depth of human serum glycan profiling.
  • This enhanced profiling capability holds potential for advancing clinical biomarker discovery through more comprehensive glycomics analysis.