Pushing the limits of native MS: Online SEC-native MS for structural biology applications
نویسندگان
چکیده
Native mass spectrometry (nMS) is now widely applied to investigate non-covalently assembled biomolecule complexes. nMS requires the use of near-neutral pH and volatile buffers preserve native state proteins. However, buffer exchange into nMS-compatible solvent usually performed manually, which results in a time-consuming tedious process, thus appearing as major drawback for analysis. Conversely, online coupling size exclusion chromatography (SEC) affords fast-automated improved desalting, but also provides an additional dimension separation complex protein mixtures. We illustrate here benefits SEC-nMS compared manual offline desalting characterization wide variety biological systems, ranging from multiprotein assemblies, protein–ligand protein–nucleic acid complexes, proteins detergent environment. then highlight potential further integrate ion mobility while preserving conformations proteins, allowing rapid collision cross section measurement even collision-induced unfolding experiments. Finally, we show that SEC can serve basis multidimensional non-denaturing liquid (LC) workflows, with acting fast device, helping achieve first LC optimal chromatographic conditions being compatible
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ژورنال
عنوان ژورنال: International Journal of Mass Spectrometry
سال: 2021
ISSN: ['1387-3806', '1873-2798']
DOI: https://doi.org/10.1016/j.ijms.2020.116502