A Liquid Chromatography/Mass Spectrometry Method for Screening Disulfide Tethering Fragments
Kenneth K Hallenbeck1, Julia L Davies1, Connie Merron1
11 Department of Pharmaceutical Chemistry and Small Molecule Discovery Center, University of California, San Francisco, San Francisco, CA, USA.
SLAS Discovery : Advancing Life Sciences R & D
|September 26, 2017
Summary
A new high-throughput liquid chromatography/mass spectrometry (LC/MS) method rapidly identifies covalent small-molecule protein binders. This optimized screening approach enhances speed and reliability for drug discovery research.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Drug Discovery
Background:
- Identifying covalent small-molecule binders to proteins is crucial for drug development.
- Existing screening methods can be time-consuming and lack flexibility for diverse targets.
Purpose of the Study:
- To refine a high-throughput liquid chromatography/mass spectrometry (LC/MS) method for identifying covalent protein binders.
- To optimize the screening process for speed, reliability, and applicability to various protein targets.
Main Methods:
- Utilized a custom library of 1600 disulfide-capped fragments targeting surface cysteine residues.
- Optimized sample preparation, chromatography, and ionization conditions for LC/MS analysis.
- Developed a custom data processing pipeline to calculate percent bound and assess signal significance.
Main Results:
- Achieved a data collection rate of 84 seconds per sample, balancing speed and reliability.
- Demonstrated method applicability across diverse protein classes and molecular masses over a 24-month period.
- Enabled screening of large covalent compound libraries within 1-2 days.
Conclusions:
- The refined LC/MS method offers a fast, reliable, and flexible platform for covalent binder screening.
- This approach significantly accelerates the identification of potential therapeutic agents targeting proteins.
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