High-Throughput Bioassays using "Dip-and-Go" Multiplexed Electrospray Mass Spectrometry

Area of Science:

• Analytical Chemistry
• Biochemistry
• Mass Spectrometry

Background:

• High-throughput screening (HTS) is crucial for drug discovery.
• Existing HTS methods can be time-consuming and complex.
• Need for faster, more efficient bioanalytical techniques.

Purpose of the Study:

• To develop a multiplexed system for HTS bioassays using inductive nanoelectrospray mass spectrometry (nESI-MS).
• To enable rapid sample loading, purification, and analysis in a 96-well microtiter plate format.
• To improve the speed and efficiency of quantitative peptide analysis in biological matrices.

Main Methods:

• Development of a multiplexed system integrating inductive nESI and micro-electrophoresis.
• Implementation of a "dip-and-go" sample loading and purification strategy.
• Automation prototype designed for parallel processing of 12 samples.

Main Results:

• Achieved efficient in situ separation and clean-up of biological samples.
• Demonstrated quantitative analysis of peptides (1-10,000 nm) in biological matrices.
• Sample processing and MS analysis achieved speeds of 20s for clean-up and 1.3-3.5s per sample for MS.

Conclusions:

• The developed nESI-MS system significantly accelerates HTS bioassays.
• Quantitative analysis of BACE1-catalyzed peptide hydrolysis showed results comparable to LC-MS but much faster.
• This technology offers a promising approach for rapid drug discovery screening.