A Hybrid Orbitrap‐Nanoelectromechanical Systems Approach for the Analysis of Individual, Intact Proteins in Real Time

• Published in: Angewandte Chemie International Edition Vol. 63; no. 33; pp. e202317064 - n/a
• Main Authors: Neumann, Adam P., Sage, Eric, Boll, Dmitri, Reinhardt‐Szyba, Maria, Fon, Warren, Masselon, Christophe, Hentz, Sébastien, Sader, John E., Makarov, Alexander, Roukes, Michael L.
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 12.08.2024
• Edition: International ed. in English
• Subjects: Chaperonin 60 - chemistry; E coli; Escherichia coli; Escherichia coli Proteins - analysis; Escherichia coli Proteins - chemistry; Hybrid systems; Instrumentation; Ion optics; Mass Spectrometry; Mass spectroscopy; Micro-Electrical-Mechanical Systems - instrumentation; Nanoelectromechanical systems; Nanomechanical systems; Nanotechnology; native mass spectrometry; Optics; Proteomics; Resonance; mass spectrometry; native mass spectrometry; Nanomechanical systems
• ISSN: 1433-7851; 1521-3773; 1521-3773
• DOI: 10.1002/anie.202317064

Nanoelectromechanical systems (NEMS)‐based mass spectrometry (MS) is an emerging technique that enables determination of the mass of individual adsorbed particles by driving nanomechanical devices at resonance and monitoring the real‐time changes in their resonance frequencies induced by each single molecule adsorption event. We incorporate NEMS into an Orbitrap mass spectrometer and report our progress towards leveraging the single‐molecule capabilities of the NEMS to enhance the dynamic range of conventional MS instrumentation and to offer new capabilities for performing deep proteomic analysis of clinically relevant samples. We use the hybrid instrument to deliver E. coli GroEL molecules (801 kDa) to the NEMS devices in their native, intact state. Custom ion optics are used to focus the beam down to 40 μm diameter with a maximum flux of 25 molecules/second. The mass spectrum obtained with NEMS‐MS shows good agreement with the known mass of GroEL. Nanoelectromechanical systems‐based mass spectrometry (NEMS‐MS) determines the mass of individual particles by monitoring resonance frequency changes in tiny mechanical devices. Incorporating NEMS into an Orbitrap instrument enhances the dynamic range and enables deep proteomic analysis. This work measures E. coli GroEL molecules, showing good agreement with the known mass.