Generation of Multiple Reporter Ions from a Single Isobaric Reagent Increases Multiplexing Capacity for Quantitative Proteomics

Craig R. Braun, Gregory H. Bird, Martin Wühr, Brian K. Erickson, Ramin Rad, Loren D. Walensky, Steven P. Gygi, Wilhelm Haas

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

Isobaric labeling strategies for mass spectrometry-based proteomics enable multiplexed simultaneous quantification of samples and therefore substantially increase the sample throughput in proteomics. However, despite these benefits, current limits to multiplexing capacity are prohibitive for large sample sizes and impose limitations on experimental design. Here, we introduce a novel mechanism for increasing the multiplexing density of isobaric reagents. We present Combinatorial Isobaric Mass Tags (CMTs), an isobaric labeling architecture with the unique ability to generate multiple series of reporter ions simultaneously. We demonstrate that utilization of multiple reporter ion series improves multiplexing capacity of CMT with respect to a commercially available isobaric labeling reagent with preserved quantitative accuracy and depth of coverage in complex mixtures. We provide a blueprint for the realization of 16-plex reagents with 1 Da spacing between reporter ions and up to 28-plex at 6 mDa spacing using only 5 heavy isotopes per reagent. We anticipate that this improvement in multiplexing capacity will further advance the application of quantitative proteomics, particularly in high-throughput screening assays.

Original languageEnglish (US)
Pages (from-to)9855-9863
Number of pages9
JournalAnalytical Chemistry
Volume87
Issue number19
DOIs
StatePublished - Oct 6 2015
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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