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Reproducibility of Differential Proteomic Technologies in CPTAC Fractionated Xenografts.

Journal of proteome research | 2016

The NCI Clinical Proteomic Tumor Analysis Consortium (CPTAC) employed a pair of reference xenograft proteomes for initial platform validation and ongoing quality control of its data collection for The Cancer Genome Atlas (TCGA) tumors. These two xenografts, representing basal and luminal-B human breast cancer, were fractionated and analyzed on six mass spectrometers in a total of 46 replicates divided between iTRAQ and label-free technologies, spanning a total of 1095 LC-MS/MS experiments. These data represent a unique opportunity to evaluate the stability of proteomic differentiation by mass spectrometry over many months of time for individual instruments or across instruments running dissimilar workflows. We evaluated iTRAQ reporter ions, label-free spectral counts, and label-free extracted ion chromatograms as strategies for data interpretation (source code is available from http://homepages.uc.edu/~wang2x7/Research.htm ). From these assessments, we found that differential genes from a single replicate were confirmed by other replicates on the same instrument from 61 to 93% of the time. When comparing across different instruments and quantitative technologies, using multiple replicates, differential genes were reproduced by other data sets from 67 to 99% of the time. Projecting gene differences to biological pathways and networks increased the degree of similarity. These overlaps send an encouraging message about the maturity of technologies for proteomic differentiation.

Pubmed ID: 26653538 RIS Download

Research resources used in this publication

None found

Antibodies used in this publication

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Associated grants

  • Agency: NCI NIH HHS, United States
    Id: U24-CA-160034
  • Agency: NCI NIH HHS, United States
    Id: U24 CA160034
  • Agency: NCI NIH HHS, United States
    Id: U24-CA-159988
  • Agency: NCI NIH HHS, United States
    Id: U24 CA159988
  • Agency: NCI NIH HHS, United States
    Id: U24-CA-160019
  • Agency: NCI NIH HHS, United States
    Id: P30 CA016086
  • Agency: NCRR NIH HHS, United States
    Id: UL1 RR024992
  • Agency: NCI NIH HHS, United States
    Id: HHSN261201100106C
  • Agency: NCI NIH HHS, United States
    Id: U24 CA160036
  • Agency: NCI NIH HHS, United States
    Id: P30 CA091842
  • Agency: NCI NIH HHS, United States
    Id: U24-CA-160036
  • Agency: NCI NIH HHS, United States
    Id: U24 CA160019
  • Agency: NCATS NIH HHS, United States
    Id: UL1 TR000448
  • Agency: NCI NIH HHS, United States
    Id: U24 CA160035
  • Agency: PHS HHS, United States
    Id: HHSN261201100106C
  • Agency: NCI NIH HHS, United States
    Id: U24-CA-160035

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