Ultrahigh-Throughput Intact Protein Analysis with Acoustic Ejection Mass Spectrometry

Area of Science:

• Biopharmaceutical analysis
• Analytical chemistry
• Mass spectrometry

Background:

• High-throughput intact protein analysis is crucial for drug discovery, particularly for protein therapeutics and covalent modifiers.
• Liquid chromatography-mass spectrometry (LC-MS) is the standard but lacks the speed for large sample sets.
• Acoustic Ejection Mass Spectrometry (AEMS) offers a faster alternative for electrospray ionization-mass spectrometry (ESI-MS) based analyses.

Purpose of the Study:

• To evaluate the application of AEMS for high-throughput intact protein analysis.
• To assess the data quality and speed of a prototype HR QTOF AEMS system for proteins (17-150 kDa).

Main Methods:

• Utilized a prototype high-resolution quadrupole time-of-flight (HR QTOF) based Acoustic Ejection Mass Spectrometry (AEMS) system.
• Analyzed intact proteins across a molecular weight range of 17 to 150 kDa.
• Compared AEMS performance against established LC-MS methods.

Main Results:

• AEMS demonstrated comparable data quality to LC-MS for intact protein analysis.
• Achieved significantly improved analytical speed, with analysis times of one second per sample.
• Successfully analyzed proteins ranging from 17 to 150 kDa.

Conclusions:

• Acoustic Ejection Mass Spectrometry (AEMS) provides an ultrahigh-throughput platform for intact protein analysis.
• This AEMS system offers a viable alternative to LC-MS, enhancing speed without compromising data quality.
• The technology holds significant potential for broad application in drug discovery and development.