High throughput screening via mass spectrometry: a case study using acetylcholinesterase

Area of Science:

• Biochemistry and Analytical Chemistry
• Drug Discovery and Development
• Enzyme Inhibition Assays

Background:

• Traditional screening methods struggle with complex substrates like lipids and steroids.
• Mass spectrometry (MS) offers label-free assay potential but is limited by low throughput.
• High-throughput screening (HTS) is crucial for efficient lead discovery in pharmaceutical research.

Purpose of the Study:

• To develop and validate a novel MS-based technique and hardware for high-throughput lead discovery.
• To enable label-free assays using native substrates for diverse enzymatic targets.
• To demonstrate the system's utility by identifying acetylcholinesterase inhibitors for Alzheimer's disease.

Main Methods:

• Development of a specialized MS system for rapid sample analysis (4-5 seconds per sample).
• Design of label-free assays utilizing biologically native substrates for enzymatic targets.
• Application of the MS system to screen a chemical library for acetylcholinesterase inhibitors.

Main Results:

• The developed MS system achieves high throughput, overcoming a major limitation of MS-based screening.
• Label-free assays were successfully implemented for direct analyte quantification in complex mixtures.
• Several potent acetylcholinesterase inhibitors were identified, with IC50 values determined.

Conclusions:

• The novel MS-based technique significantly enhances throughput for lead discovery applications.
• This approach is versatile, applicable to a broad range of enzymatic targets and complex substrates.
• The system demonstrates efficacy in identifying clinically relevant inhibitors, such as those for Alzheimer's disease.