Protein identification by 3D OrbiSIMS to facilitate in situ imaging and depth profiling

Anna M. Kotowska, Gustavo F. Trindade, Paula M. Mendes, Philip M. Williams, Jonathan W. Aylott, Alexander G. Shard, Morgan R. Alexander and David J. Scurr ()
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Anna M. Kotowska: University of Nottingham
Gustavo F. Trindade: University of Nottingham
Paula M. Mendes: University of Birmingham
Philip M. Williams: University of Nottingham
Jonathan W. Aylott: University of Nottingham
Alexander G. Shard: National Physical Laboratory
Morgan R. Alexander: University of Nottingham
David J. Scurr: University of Nottingham

Nature Communications, 2020, vol. 11, issue 1, 1-8

Abstract: Abstract Label-free protein characterization at surfaces is commonly achieved using digestion and/or matrix application prior to mass spectrometry. We report the assignment of undigested proteins at surfaces in situ using secondary ion mass spectrometry (SIMS). Ballistic fragmentation of proteins induced by a gas cluster ion beam (GCIB) leads to peptide cleavage producing fragments for subsequent OrbitrapTM analysis. In this work we annotate 16 example proteins (up to 272 kDa) by de novo peptide sequencing and illustrate the advantages of this approach by characterizing a protein monolayer biochip and the depth distribution of proteins in human skin.

Date: 2020
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DOI: 10.1038/s41467-020-19445-x

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