Related Experiment Video
Updated: May 19, 2026
09:18
Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics
Published on: April 17, 2017
Integrating Simple Microfluidics Design/Fabrication with a Novel Ionization Source for Time-Resolved Chemical
Amanda Overby1, Toktam Godary1, Sydney Anderson1
1Department of Chemistry, West Virginia University, Morgantown, West Virginia 26506, United States.
ACS Omega
|May 18, 2026
Summary
A novel 3D-printed microfluidic device enhances mixing for time-resolved hydrogen-deuterium exchange (HDX) mass spectrometry (MS) experiments. This innovation enables precise analysis of biomolecular dynamics with improved efficiency and reduced noise.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Chemical Engineering
Background:
- Time-resolved hydrogen-deuterium exchange (HDX) coupled with mass spectrometry (MS) is crucial for studying protein dynamics.
- Efficient mixing of reagents is a key challenge in achieving high temporal resolution in HDX-MS.
- Existing microfluidic devices may face limitations in mixing efficiency and flow rate versatility.
Purpose of the Study:
- To develop and validate a novel 3D-printed microfluidic device for time-resolved HDX-MS.
- To integrate efficient mixing with high-efficiency capillary vibrating sharp-edge spray ionization (cVSSI) for enhanced analytical performance.
- To demonstrate the device's capability in studying biomolecular dynamics across a wide range of experimental conditions.
Main Methods:
- Fabrication of a 3D-printed microfluidic device with a dual herringbone mixer and three inlet ports.
- Integration of the microfluidic device with a capillary vibrating sharp-edge spray ionization (cVSSI) source and mass spectrometer (MS).
- Performance evaluation using time-resolved hydrogen-deuterium exchange experiments on the model protein myoglobin.
Main Results:
- The 3D-printed device achieved high-efficiency mixing of reagent streams over a >10-fold range of flow rates.
- The integrated cVSSI-MS system demonstrated operability across a wide flow rate range suitable for time-dependent reactions.
- Reproducible HDX measurements were obtained for myoglobin, showcasing the device's potential for biomolecular studies.
Conclusions:
- The developed 3D-printed microfluidic device coupled with cVSSI-MS provides a robust platform for time-resolved HDX analysis.
- The omniphobic surface rendering effectively reduced chemical noise during ionization.
- This early-stage device offers a promising tool for advancing the study of protein dynamics and other time-dependent molecular interactions.
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