Development of Modular High-Resolution Ion Mobility Orbitrap Platforms for Native Mass Spectrometry

Abstract

Native ion mobility-mass spectrometry (IM-MS) has evolved into a multifunctional tool for the characterization and analysis of native intact protein complexes. Structural information can be inferred by the rotationally-averaged collision cross-section (CCS) measurements. Native IM-MS is complementary to nuclear magnetic resonance (NMR) spectroscopy, X-Ray crystallography (XRD), cryogenic-electron microscopy (CryoEM), as well as surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC). Commercial IM-MS platforms excel with proteomic workflows but are limited in resolution, sensitivity, and mass range for large biomolecules. Structural biology problems require developing and applying new instruments that excel in the sensitivity, resolution, and resolving power of large biological systems (MW > 100 kDa), e.g., proteins, protein complexes, antibodies, chaperonins, and virus capsids. The work described vide infra focuses on developing and implementing novel home-built instrumentation for native IM-MS to expand the capabilities of the biophysical toolbox in structural biology.