Voltage Rollercoaster Filtering of Low-Mass Contaminants During Native Protein Analysis

John P. McGee, Rafael D. Melani, Michael Goodwin, Graeme McAlister, Romain Huguet, Michael W. Senko, Philip D. Compton, Neil L. Kelleher

Research output: Contribution to journalArticle

1 Scopus citations

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 languageEnglish (US)
Pages (from-to)763-767
Number of pages5
JournalJournal of the American Society for Mass Spectrometry
Volume31
Issue number3
DOIs
StatePublished - Mar 4 2020

ASJC Scopus subject areas

  • Structural Biology
  • Spectroscopy

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