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11:32
A Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) Platform for Investigating Peptide Biosynthetic Enzymes
Published on: May 4, 2020
CE hydrogen deuterium exchange-MS in peptide analysis
Simon S M Lau1, Nick M Stainforth, David G Watson
1Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, UK.
Electrophoresis
|December 15, 2007
Summary
This study integrates capillary electrophoresis (CE) with hydrogen-deuterium (H/D) exchange mass spectrometry (MS) for peptide analysis. The combined method provides clear mass spectra for accurate amino acid composition and sequence confirmation.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Mass Spectrometry
Background:
- Capillary electrophoresis (CE) and hydrogen-deuterium (H/D) exchange mass spectrometry (MS) are established techniques for peptide analysis.
- Integrating these methods offers potential for enhanced characterization of peptides.
Purpose of the Study:
- To report the facile coupling of CE and H/D exchange MS for analyzing model and pharmaceutical peptides.
- To demonstrate the utility of this combined approach for peptide characterization.
Main Methods:
- Utilized a sheath flow interface for coupling CE with H/D exchange MS.
- Employed specific buffer compositions in D(2)O and solvent mixtures to facilitate complete H/D exchange.
- Analyzed peptides with varying masses and charge states, including leucine enkephalin, bombesin, and substance P.
Main Results:
- Achieved complete H/D exchange for peptides ranging from 556 to 1620 Da.
- Generated easily interpretable ESI mass spectra comparable to LC-MS.
- Observed predicted b- and y-series fragments from in-source decomposition.
- Demonstrated confirmation of amino acid composition and sequence through H/D exchange on molecular and fragment ions.
Conclusions:
- The CE-H/D-MS approach is effective for complete H/D exchange in peptide analysis.
- This method requires lower volumes of deuterated solvents compared to other techniques.
- The technique aids in confirming peptide amino acid composition and sequence.
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