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Implementation of Intact Protein MS on Commercial SLIM Instrumentation using nano-, micro-, and high- flow ionization sources and applications to disease associated proteoforms and intact antibodies
  • +7
  • Md Amin Hossain,
  • Tom Doherty,
  • Jennifer Krone,
  • Daniel DeBord,
  • Luis Viskatis,
  • Jared R. Auclair,
  • Thais Pedrete,
  • Fanny Liu,
  • Christian Bleiholder,
  • Jeff Agar
Md Amin Hossain
Northeastern University Department of Chemistry and Chemical Biology
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Tom Doherty
MOBILion Systems Inc
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Jennifer Krone
MOBILion Systems Inc
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Daniel DeBord
MOBILion Systems Inc
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Luis Viskatis
Northeastern University Department of Chemistry and Chemical Biology
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Jared R. Auclair
Northeastern University Department of Chemistry and Chemical Biology
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Thais Pedrete
Florida State University
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Fanny Liu
Florida State University
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Christian Bleiholder
Florida State University
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Jeff Agar
Northeastern University Department of Chemistry and Chemical Biology

Corresponding Author:[email protected]

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Abstract

This study addresses three unmet needs: 1) development of widely accessible native mass spectrometry (MS) methods and 2) native top-down mass spectrometry (TDMS) methods for high flow sources on an Agilent Q-TOF mass spectrometer, and 3) confirming suitability of structures for lossless ion manipulation (SLIM) ion mobility (IM) MS methods for analysis of native intact proteins. To decrease the cost and training barrier to performing MS, TDMS, and IM-MS analysis of native proteins, we have developed methods for nanospray, microspray, and heated high flow electrospray ionization sources. SLIM offers the highest full-scan IM resolution available, but commercial instrumentation has not previously been applied to native MS. Our optimized methods extend the upper m/z limit beyond the manufacturer’s default IM mode upper mass limit of 4000 m/z to 7000 m/z, which allows for intact protein and native MS analysis of a variety of proteins, including native MS of intact IgG antibodies. Using the robust and facile Agilent Jet Spray (AJS) source, we demonstrate the application of native protein MS to amyotrophic lateral sclerosis (ALS)-associated Cu/Zn-superoxide dismutase proteoforms, characterizing a novel conformer and quaternary structural changes.
Submitted to Rapid Communications in Mass Spectrometry
16 Jun 2024Reviewer(s) Assigned
12 Jul 2024Review(s) Completed, Editorial Evaluation Pending
23 Jul 2024Editorial Decision: Revise Major