Multiple Reaction Monitoring (MRM) is routinely used for protein biomarker qualification, verification and validation in large clinical cohorts. A typical study may include the monitoring of several hundreds of peptides across hundreds or thousands of plasma/serum samples. Peak integration of these large scale data sets is both very challenging and time consuming, due to the need for manual inspection and correction of incorrectly integrated peaks. This presentation will focus on the development of a high-throughput peak integration workflow for the analysis of large-scale proteomics datasets using Skyline and in-house developed tools. Datasets containing several hundreds of thousands of peaks can be integrated and QC’d within 2 days with a minimal amount of manual inspection. This same workflow is also used for the automatic monitoring of instrument performance (sensitivity, RT reproducibility and peak width of target peptides) during the analysis of the study samples.