Analysis of Proteins, Protein Complexes, and Organellar Proteomes Using Sheathless Capillary Zone Electrophoresis - Native Mass Spectrometry

• DOI: 10.1007/s13361-017-1781-1
• Publication Date: 2017-09-05T13:47:38Z
• Authors: Arseniy M. Belov; Rosa Viner; Marcia R. Santos; David M. Horn; Marshall Bern; Barry L. Karger; Alexander R. Ivanov
• Keywords: Orbitrap; Proteome; Sample Preparation

Native mass spectrometry (MS) is a rapidly advancing field in the analysis of proteins, protein complexes, and macromolecular species various types. The majority native MS experiments reported to-date has been conducted using direct infusion purified analytes into spectrometer. In this study, capillary zone electrophoresis (CZE) was coupled online to Orbitrap spectrometers commercial sheathless interface enable high-performance separation, identification, structural characterization limited amounts proteins latter with preserved non-covalent associations under conditions. performance both bare-fused silica polyacrylamide-coated capillaries assessed mixtures standards known form protein-protein protein-ligand complexes. High-efficiency separation complexes demonstrated types, while polyacrylamide neutral-coated showed better reproducibility higher efficiency for more complex samples. platform then evaluated determination monoclonal antibody aggregation proteomes complexity ribosomal isolate from E. coli. CZE-MS, accurate single stage tandem-MS measurements, enabled identification proteoforms at femtomole levels. This study demonstrates that CZE-MS can serve as an orthogonal complementary technique conventional methodologies advantages low sample consumption, minimal processing losses, high throughput sensitivity. presents novel ribosomes other organelles, potential discovery features defining cellular phenotypes (e.g., specialized ribosomes). Graphical Abstract ᅟ.