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Resolving heterogeneous high-mass macromolecular machineries by Orbitrap-based single particle charge detection mass spectrometry

Tobias P. Wörner, Joost Snijder, Antonette Bennett, Mavis Agbandje-McKenna, Alexander A. Makarov, Albert J.R. Heck
doi: https://doi.org/10.1101/717413
Tobias P. Wörner
1Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands
2Netherlands Proteomics Center, Padualaan 8, 3584 CH Utrecht, The Netherlands
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Joost Snijder
1Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands
2Netherlands Proteomics Center, Padualaan 8, 3584 CH Utrecht, The Netherlands
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Antonette Bennett
3Department of Biochemistry and Molecular Biology, Center for Structural Biology, the McKnight Brain Institute, 1200 Newell Drive, Gainesville, FL 32610, USA
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Mavis Agbandje-McKenna
3Department of Biochemistry and Molecular Biology, Center for Structural Biology, the McKnight Brain Institute, 1200 Newell Drive, Gainesville, FL 32610, USA
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Alexander A. Makarov
1Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands
4Thermo Fisher Scientific (Bremen), Bremen, Germany
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Albert J.R. Heck
1Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands
2Netherlands Proteomics Center, Padualaan 8, 3584 CH Utrecht, The Netherlands
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  • For correspondence: a.j.r.heck@uu.nl
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Abstract

Here we show that single particle charge-detection mass spectrometry (CD-MS) can be performed on a ubiquitous Orbitrap mass analyser and applied to the analysis of high-mass (megadalton) heterogeneous biomolecular assemblies. We demonstrate that single particle high-mass ions can survive in the Orbitrap for seconds, whereby their measured signal amplitudes scale linearly with charge state over the entire m/z range. Orbitrap based single particle CD-MS can be used to resolve mixed ion populations, accurately predict charge states, and consequently also the mass of the ions. We successfully applied CD-MS to challenging natural and biotherapeutic protein assemblies, such as IgM oligomers, designed protein nano-cages, ribosome particles and intact, empty- and genome-loaded Adeno-associated virus particles. Single particle CD-MS combined with native MS on existing Orbitrap platforms will greatly expand its application, especially in the mass analysis of megadalton heterogeneous biomolecular assemblies.

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Posted July 28, 2019.
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Resolving heterogeneous high-mass macromolecular machineries by Orbitrap-based single particle charge detection mass spectrometry
Tobias P. Wörner, Joost Snijder, Antonette Bennett, Mavis Agbandje-McKenna, Alexander A. Makarov, Albert J.R. Heck
bioRxiv 717413; doi: https://doi.org/10.1101/717413
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Resolving heterogeneous high-mass macromolecular machineries by Orbitrap-based single particle charge detection mass spectrometry
Tobias P. Wörner, Joost Snijder, Antonette Bennett, Mavis Agbandje-McKenna, Alexander A. Makarov, Albert J.R. Heck
bioRxiv 717413; doi: https://doi.org/10.1101/717413

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