Achieving a 35-Plex Tandem Mass Tag Reagent Set through Deuterium Incorporation.

ABSTRACT: Mass spectrometry-based sample multiplexing with isobaric tags permits the development of high-throughput and precise quantitative biological assays with proteome-wide coverage and minimal missing values. Here, we nearly doubled the multiplexing capability of the TMTpro reagent set to a 35-plex through the incorporation of one deuterium isotope into the reporter group. Substituting deuterium frequently results in suboptimal peak coelution, which can compromise the accuracy of reporter ion-based quantification. To counteract the deuterium effect on quantitation, we implemented a strategy that necessitated the segregation of nondeuterium and deuterium-containing channels into distinct subplexes during normalization procedures, with reassembly through a common bridge channel. This multiplexin

SUBMITTER: Zuniga NR 

PROVIDER: S-EPMC12706455 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature