An Ultra-High Throughput Hydrogen-Deuterium Exchange Workflow Using Acoustic Ejection Mass Spectrometry for Studying...

Area of Science:

• Pharmaceutical Science
• Analytical Chemistry
• Biophysical Chemistry

Background:

• Peptides are crucial in pharmaceuticals due to their potency and specificity.
• Peptide conformation in solution influences drug properties and requires structure-based optimization.
• Current methods for studying peptide conformation are limited in throughput and resources for early drug discovery.

Purpose of the Study:

• To develop a high-throughput, sample-preparation-free workflow for peptide solution conformation analysis.
• To meet the analytical demands of large-scale screening in early drug discovery pipelines.
• To enable efficient investigation of peptide conformational dynamics.

Main Methods:

• Utilized hydrogen-deuterium exchange mass spectrometry (HDX-MS).
• Integrated HDX-MS with ultra-high throughput acoustic ejection mass spectrometry (uHT AEMS).
• Achieved a sampling rate of approximately 3 seconds per sample with minimal sample and solvent usage.

Main Results:

• Demonstrated ultra-high throughput (uHT) analysis of peptide solution conformation.
• Established the workflow's capability to discriminate conformational changes using myoglobin and apo-myoglobin models.
• Validated the method's effectiveness in differentiating solution conformations of model peptides via HDX profiles.

Conclusions:

• The developed workflow provides a high-throughput, sample-preparation-free approach for peptide conformation studies.
• uHT AEMS combined with HDX-MS is suitable for large-scale screening in early drug discovery.
• This methodology supports efficient down-selection of peptide drug leads based on solution conformation.