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

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 Orbitrap^TM 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.

SUBMITTER: Kotowska AM 

PROVIDER: S-EPMC7672064 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature