Software for quantitative proteomic analysis using stable isotope labeling and data independent acquisition

Xin Huang, Miao Liu, Michael J. Nold, Changhai Tian, Kai Fu, Jialin Zheng, Scott J. Geromanos, Shi Jian Ding

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

Many software tools have been developed for analyzing stable isotope labeling (SIL)-based quantitative proteomic data using data dependent acquisition (DDA). However, programs for analyzing SIL-based quantitative proteomics data obtained with data independent acquisition (DIA) have yet to be reported. Here, we demonstrated the development of a new software for analyzing SIL data using the DIA method. Performance of the DIA on SYNAPT G2MS was evaluated using SIL-labeled complex proteome mixtures with known heavy/light ratios (H/L = 1:1, 1:5, and 1:10) and compared with the DDA on linear ion trap (LTQ)-Orbitrap MS. The DIA displays relatively high quantitation accuracy for peptides cross all intensity regions, while the DDA shows an intensity dependent distribution of H/L ratios. For the three proteome mixtures, the number of detected SIL-peptide pairs and dynamic range of protein intensities using DIA drop stepwise, whereas no significant changes in these aspects using DDA were observed. The new software was applied to investigate the proteome difference between mouse embryonic fibroblasts (MEFs) and MEF-derived induced pluripotent stem cells (iPSCs) using 16O/ 18O labeling. Our study expanded the capacities of our UNiquant software pipeline and provided valuable insight into the performance of the two cutting-edge MS platforms for SIL-based quantitative proteomic analysis today.

Original languageEnglish (US)
Pages (from-to)6971-6979
Number of pages9
JournalAnalytical chemistry
Volume83
Issue number18
DOIs
StatePublished - Sep 15 2011

ASJC Scopus subject areas

  • Analytical Chemistry

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