Expanding Differential Ion Mobility Separations into the MegaDalton Range

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Authors
Wörner, Tobias P.
Thurman, Hayden A.
Makarov, Alexander A.
Shvartsburg, Alexandre A.
Issue Date
2023
Type
Article
Language
en_US
Keywords
Electrospray ionization , Ion mobility spectrometers , Mass spectrometry , Monoclonal antibodies , Viruses , Analytical method , Electron capture dissociation , Electron transfer dissociation , Electrospray ionization mass spectrometry , High definition , Ion Mobility , Large biomolecules , Mass spectrometry analysis , Native mass spectrometries , Protein complexes , Ions
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Abstract

Along with mass spectrometry (MS), ion mobility separations (IMS) are advancing to ever larger biomolecules. The emergence of electrospray ionization (ESI) and native MS enabled the IMS/MS analyses of proteins up to ∼100 kDa in the 1990s and whole protein complexes and viruses up to ∼10 MDa since the 2000s. Differential IMS (FAIMS) is substantially orthogonal to linear IMS based on absolute mobility K and offers exceptional resolution, unique selectivity, and steady filtering readily compatible with slower analytical methods such as electron capture or transfer dissociation (ECD/ETD). However, the associated MS stages had limited FAIMS to ions with m/z < 8000 and masses under ∼300 kDa. Here, we integrate high-definition FAIMS with the Q-Exactive Orbitrap UHMR mass spectrometer that can handle m/z up to 80,000 and MDa-size ions in the native ESI regime. In the initial evaluation, the oligomers of monoclonal antibody adalimumab (148 kDa) are size-selected up to at least the nonamers (1.34 MDa) with m/z values up to ∼17,000. This demonstrates the survival and efficient separation of noncovalent MDa assemblies in the FAIMS process, opening the door to novel analyses of the heaviest macromolecules. © 2024 American Chemical Society.

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Wörner, T.P., Thurman, H.A., Makarov, A.A., Shvartsburg, A.A. Expanding Differential Ion Mobility Separations into the MegaDalton Range. (2023). Analytical Chemistry. DOI: 10.1021/acs.analchem.3c05012
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American Chemical Society
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Analytical Chemistry
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0003-2700
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