Optimization of protein-level tandem mass tag (TMT) labeling conditions in complex samples with top-down proteomics.

ABSTRACT: Isobaric chemical tag labels (e.g., iTRAQ and TMT) have been extensively utilized as a standard quantification approach in bottom-up proteomics, which provides high multiplexing capacity and enables MS2-level quantification while not complicating the MS1 scans. We recently demonstrated the feasibility of intact protein TMT labeling for the identification and quantification with top-down proteomics of smaller intact proteoforms (<35 kDa) in complex biological samples through the removal of large proteins prior to labeling. Still, the production of side products during TMT labeling (i.e., incomplete labeling or labeling of unintended residues) complicated the analysis of complex protein samples. In this study, we systematically evaluated the protein-level TMT labeling reaction parameters, in

SUBMITTER: Guo Y 

PROVIDER: S-EPMC9371347 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature