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
Author Information
John P McGee: Departments of Chemical and Biological Engineering, Chemistry, and Molecular Biosciences, the Chemistry of Life Processes Institute, the Proteomics Center of Excellence at Northwestern University, Evanston, Illinois 60208, United States. ORCID
Rafael D Melani: Departments of Chemical and Biological Engineering, Chemistry, and Molecular Biosciences, the Chemistry of Life Processes Institute, the Proteomics Center of Excellence at Northwestern University, Evanston, Illinois 60208, United States.
Michael Goodwin: Thermo Fisher Scientific, San Jose, California 95134, United States.
Graeme McAlister: Thermo Fisher Scientific, San Jose, California 95134, United States.
Romain Huguet: Thermo Fisher Scientific, San Jose, California 95134, United States.
Michael W Senko: Thermo Fisher Scientific, San Jose, California 95134, United States.
Philip D Compton: Departments of Chemical and Biological Engineering, Chemistry, and Molecular Biosciences, the Chemistry of Life Processes Institute, the Proteomics Center of Excellence at Northwestern University, Evanston, Illinois 60208, United States. ORCID
Neil L Kelleher: Departments of Chemical and Biological Engineering, Chemistry, and Molecular Biosciences, the Chemistry of Life Processes Institute, the Proteomics Center of Excellence at Northwestern University, Evanston, Illinois 60208, United States. ORCID
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 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.
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