Abstract
Intact protein mass spectrometry (MS) via electrospray-based methods is often degraded by low-mass contaminants, which can suppress the spectral quality of the analyte of interest via space-charge effects. Consequently, selective removal of contaminants by their mobilities would benefit native MS if achieved without additional hardware and before the mass analyzer regions used for selection, analyte readout, or tandem MS. Here, we use the high-pressure multipole within the source of an Orbitrap Tribrid as the foundation for a coarse ion filter. Using this method, we show complete filtration of 2 mM polyethylene glycol (PEG-1000) during native MS of SILu mAb antibody present at a 200× lower concentration. We also show the generality of the process by rescuing 10 μM tetrameric pyruvate kinase from 2 mM PEG-1000, asserting this voltage rollercoaster filtering (VRF) method for use in native MS as an efficient alternative to conventional purification methods.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 763-767 |
| Number of pages | 5 |
| Journal | Journal of the American Society for Mass Spectrometry |
| Volume | 31 |
| Issue number | 3 |
| DOIs | |
| State | Published - Mar 4 2020 |
Funding
This research was supported by the National Resource for Translational and Developmental Proteomics under Grant P41 GM108569 from the National Institute of General Medical Sciences of the National Institutes of Health and also supported by both the Sherman Fairchild Foundation and Thermo Fisher Scientific. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The authors thank Paul Thomas, Robert Gerbasi, Jared Kafader, Kevin Jooss, Steven Patrie, and Haemin Park for their assistance in proofreading this manuscript.
ASJC Scopus subject areas
- Structural Biology
- Spectroscopy