RT Journal Article
SR Electronic
T1 Resolving heterogeneous high-mass macromolecular machineries by Orbitrap-based single particle charge detection mass spectrometry
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 717413
DO 10.1101/717413
A1 Tobias P. Wörner
A1 Joost Snijder
A1 Antonette Bennett
A1 Mavis Agbandje-McKenna
A1 Alexander A. Makarov
A1 Albert J.R. Heck
YR 2019
UL http://biorxiv.org/content/early/2019/07/27/717413.abstract
AB 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.