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Journal ArticleDOI

mProphet: automated data processing and statistical validation for large-scale SRM experiments

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TLDR
In this article, the authors present mProphet, a fully automated system that computes accurate error rates for the identification of targeted peptides in SRM data sets and maximizes specificity and sensitivity by combining relevant features in the data into a statistical model.
Abstract
Selected reaction monitoring (SRM) is a targeted mass spectrometric method that is increasingly used in proteomics for the detection and quantification of sets of preselected proteins at high sensitivity, reproducibility and accuracy. Currently, data from SRM measurements are mostly evaluated subjectively by manual inspection on the basis of ad hoc criteria, precluding the consistent analysis of different data sets and an objective assessment of their error rates. Here we present mProphet, a fully automated system that computes accurate error rates for the identification of targeted peptides in SRM data sets and maximizes specificity and sensitivity by combining relevant features in the data into a statistical model.

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Journal ArticleDOI

Systems proteomics of liver mitochondria function

TL;DR: The characterization of mitochondrial function in a genetic reference panel of recombinant inbred mice showed a frequent lack of correlation of transcript and protein abundance, enabled the identification of genomic variants of mitochondrial enzymes that caused inborn errors in metabolism, and revealed two genes that appear to function in cholesterol metabolism.
Journal ArticleDOI

A review on mass spectrometry-based quantitative proteomics: Targeted and data independent acquisition.

TL;DR: A step-by-step tutorial on development of SRM/MRM protein assay intended for researchers without prior experience in proteomics is presented.
Journal ArticleDOI

Reproducible quantification of cancer-associated proteins in body fluids using targeted proteomics

TL;DR: A high-throughput method for the development and refinement of selected reaction monitoring (SRM) assays for human proteins is described and it is demonstrated that these SRM assays allow reproducible quantification by monitoring 34 biomarker candidates across 83 patient plasma samples.
Journal ArticleDOI

Quantitative measurements of N-linked glycoproteins in human plasma by SWATH-MS.

TL;DR: Overall, SWATH‐MS measurements showed a slightly lower sensitivity and a comparable reproducibility to state‐of‐the‐art SRM measurements for targeted quantification of the N‐glycosites in human blood, but a significantly larger number of peptides can be quantified per analysis.
References
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Journal ArticleDOI

Statistical significance for genomewide studies

TL;DR: This work proposes an approach to measuring statistical significance in genomewide studies based on the concept of the false discovery rate, which offers a sensible balance between the number of true and false positives that is automatically calibrated and easily interpreted.
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Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics.

TL;DR: SILAC is a simple, inexpensive, and accurate procedure that can be used as a quantitative proteomic approach in any cell culture system and is applied to the relative quantitation of changes in protein expression during the process of muscle cell differentiation.
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Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search.

TL;DR: A statistical model is presented to estimate the accuracy of peptide assignments to tandem mass (MS/MS) spectra made by database search applications such as SEQUEST, demonstrating that the computed probabilities are accurate and have high power to discriminate between correctly and incorrectly assigned peptides.
Journal ArticleDOI

A statistical model for identifying proteins by tandem mass spectrometry.

TL;DR: A statistical model is presented for computing probabilities that proteins are present in a sample on the basis of peptides assigned to tandem mass (MS/MS) spectra acquired from a proteolytic digest of the sample, and it is shown to produce probabilities that are accurate and have high power to discriminate correct from incorrect protein identifications.
Journal ArticleDOI

Skyline: an open source document editor for creating and analyzing targeted proteomics experiments

TL;DR: The Skyline user interface simplifies the development of mass spectrometer methods and the analysis of data from targeted proteomics experiments performed using selected reaction monitoring (SRM).
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