Open AccessVoltage Rollercoaster Filtering of Low-Mass Contaminants During Native Protein Analysis
Author(s) -
John P. McGee,
Rafael D. Melani,
Michael Goodwin,
Graeme C. McAlister,
Romain Huguet,
Michael W. Senko,
Philip D. Compton,
Neil L. Kelleher
Publication year - 2020
Publication title -
journal of the american society for mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.961
H-Index - 127
eISSN - 1879-1123
pISSN - 1044-0305
DOI - 10.1021/jasms.9b00037
Subject(s) - chemistry , analyte , mass spectrometry , chromatography , electrospray , analytical chemistry (journal) , polyethylene glycol , filter (signal processing) , organic chemistry , computer science , computer vision
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.