Analyzing LC-MS/MS data by spectral count and ion abundance: two case studies

TL;DR

This study compares spectral count and ion abundance measures in LC-MS/MS proteomics, demonstrating ion abundance's higher sensitivity and how data aggregation methods influence analysis conclusions.

Contribution

It provides a comparative analysis of spectral count and ion abundance, highlighting ion abundance's superior sensitivity and the impact of data rollup methods on results.

Findings

Ion abundance is more sensitive than spectral count.

Different data rollup methods can lead to divergent conclusions.

Ion abundance reveals properties not seen with spectral count.

Abstract

In comparative proteomics studies, LC-MS/MS data is generally quantified using one or both of two measures: the spectral count, derived from the identification of MS/MS spectra, or some measure of ion abundance derived from the LC-MS data. Here we contrast the performance of these measures and show that ion abundance is the more sensitive. We also examine how the conclusions of a comparative analysis are influenced by the manner in which the LC-MS/MS data is `rolled up' to the protein level, and show that divergent conclusions obtained using different rollups can be informative. Our analysis is based on two publicly available reference data sets, BIATECH-54 and CPTAC, which were developed for the purpose of assessing methods used in label-free differential proteomic studies. We find that the use of the ion abundance measure reveals properties of both data sets not readily apparent using the spectral count.