Direct Analysis from Phase-Separated Liquid Samples using ADE-OPI-MS: Applicability to High-Throughput Screening for...

Area of Science:

• Biochemistry
• Analytical Chemistry
• Pharmacology

Background:

• High-throughput screening (HTS) identifies compounds modulating therapeutic targets.
• Mass spectrometry (MS) offers label-free detection for HTS, overcoming optical technique limitations.
• Conventional MS platforms (e.g., LC-MS) have throughput limitations for large compound libraries.

Purpose of the Study:

• To characterize an acoustic droplet ejection, open port interface-mass spectrometry (ADE-OPI-MS) system for HTS.
• To assess the ADE-OPI-MS platform's suitability for screening membrane-associated enzymes.
• To evaluate the efficiency and performance of ADE-OPI-MS compared to existing HTS methods.

Main Methods:

• Development and characterization of an ADE-OPI-MS system.
• Utilizing diacylglycerol acyltransferase 2 (DGAT2) as a model enzyme system.
• Screening a 6400-member compound library using the ADE-OPI-MS platform.

Main Results:

• The ADE-OPI-MS system successfully ejected droplets from phase-separated samples, enabling direct coupling of liquid-liquid extraction with OPI-MS.
• The platform demonstrated suitability for HTS applications.
• ADE-OPI-MS screening performance was comparable to LC-MS, achieving over 20-fold greater efficiency.

Conclusions:

• The ADE-OPI-MS system represents a significant advancement in chromatography-free MS-based HTS.
• This novel platform enhances screening efficiency for drug discovery campaigns.
• The system's ability to handle phase-separated samples broadens its applicability in biochemical assays.