Overview

Highlights

  • Intensity-based quantification with high accuracy
  • Works for both label-free and labeling data, including:
    • SILAC, iTRAQ, TMT(MS2/MS3), N-Terminal
  • Support complex experimental design
  • Integrated with database search

Stable isotope labeling using amino acids in cell culture (SILAC) is a powerful method to identify and quantify relative differential changes in complex protein samples. It has become the most common approach for in vivo isotopic labeling and popular method for quantitative proteomics.
The PEAKS Q module in PEAKS Studio 8.5 adopts
1) enhanced SILAC feature pair detection algorithm, and
2) SILAC alignment and ID transfer techniques to successfully improve the quantification accuracy and sensitivity. It uses ratio-focused quantification, which enables the data analysis for super-SILAC experiments.

 

It enables applications such as time series quantification by introducing multiple group comparison. This new PEAKS Q module also implements paired t-test and Welch’s ANOVO for single-group analysis and multi-group comparison, respectively.
Protein relative quantification in PEAKS is performed using ion peak intensity on MS1. In label-free quantification experiments, samples are separately collected, prepared and analyzed by LC-MS/MS. Because of the large amount of data collected from these experiments, sensitive and accurate algorithms are used in PEAKS for automated ion peak alignment and comparison.
Peptide/protein identification from MS2 by database search is integrated for protein quantification.
PEAKS Studio 8.5 selects the three most abundant unique peptides for protein ratio estimation by excluding

 

1) Peptides with both modified and unmodified forms, and
2) Redundant peptides.
Different approaches to the t-test or ANOVA are required for robustly comparing pairs of independent or related samples. When a large number of tests are performed, P values are interpreted differently for more accurate estimation of false discovery rate (FDR).
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Isobaric tags (TMT or iTRAQ) have identical masses and chemical properties that allow heavy and light isotopologues to co-elute together. The tags are then cleaved from the peptides by collision-induced dissociation (CID) during MS/MS, which is used for quantification.
One of the challenges for such methods is reporter ion ratio distortion resulting from fragmentation of co-isolated interfering species.

 

The MultiNotch MS3 method addresses this issue by uniquely combining multiplexing capacity with quantitative sensitivity and accuracy. PEAKS supports both MS2 and MS3 quantification.
With PEAKS, you can now expand your sample size for large-scale protein quantification studies, with reference channels to ensure the accuracy of quantification.
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