The analysis of data-independent acquisition (DIA) mass spectrometry data is crucial for comprehensive proteomics studies. However, traditional single-run methods often fall short in terms of identification depth and consistency. We present HFDiscrim, a specialized multirun DIA analysis tool aimed at enhancing the depth and consistency of reliable peptide identifications of DIA analysis tools. HFDiscrim was extensively benchmarked on multiple data sets, including the MCB data set, the ccRCC data set, and a three-species benchmark mixture. Compared to PyProphet, HFDiscrim identified 22.04% more precursors, 19.1% more peptides, and 13.2% more proteins while maintaining a controllable false discovery rate. Furthermore, HFDiscrim demonstrated higher identification rates and improved reproducibility across multiple runs. HFDiscrim is publicly available at https://github.com/yachliu/HFDiscrim.
© 2024 The Authors. Published by American Chemical Society.