Ultrahigh-Throughput Intact Protein Analysis with Acoustic Ejection Mass Spectrometry

J Am Soc Mass Spectrom. 2023 Jan 4;34(1):4-9. doi: 10.1021/jasms.2c00276. Epub 2022 Dec 5.

Abstract

The need for high-throughput intact protein analysis has been rising as drug discovery increasingly requires the analysis of large sets of covalent modifiers and protein therapeutics. Liquid chromatography-mass spectrometry (LC-MS) is the primary analytical tool used to date to characterize proteins within the biopharmaceutical industry. However, the speed of LC-MS prevents the analysis of large-scale sample sets (>1000 within a day). Acoustic ejection mass spectrometry (AEMS) has recently been established as an electrospray ionization (ESI)-MS based platform with both fast analytical throughput and high data quality. Since its introduction, this technology has been applied in numerous fields with a primary focus on small-molecule analysis in high-throughput drug discovery and development. Here we explore the application of AEMS to high-throughput intact protein analysis for proteins ranging in molecular weight from 17 to 150 kDa on a prototype high-resolution quadrupole time-of-flight (HR QTOF) based AEMS system. Data quality obtained on this platform is comparable to LC-MS, while the analysis speed is significantly improved to one-second-per-sample. This ultrahigh-throughput intact protein analysis platform has the potential to be used broadly in drug discovery.

MeSH terms

  • Acoustics
  • Chromatography, Liquid / methods
  • Mass Spectrometry / methods
  • Proteins* / chemistry
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Sulfones*

Substances

  • 2-aminoethylmethyl sulfone
  • Sulfones
  • Proteins