Scoring Information Integration with Statistical Quality Control Enhanced Cross-Run Analysis of Data-Independent Acquisition Proteomics Data

The peptide-centric strategy is widely applied in data-independent acquisition (DIA) proteomics to analyze multiplexed MS2 spectra. However, current software tools often rely on single-run data for peptide peak identification, leading to inconsistent quantification across heterogeneous datasets. Match-between-runs (MBR) algorithms address this by aligning peaks or elution profiles across runs post-analysis but they are often ad-hoc and lack statistical frameworks for controlling peak quality, resulting in false positives and reduced quantitative reproducibility. Here we present DreamDIAlignR, a cross-run peptide-centric tool that integrates peptide elution behavior across runs with a deep learning peak identifier and signal alignment algorithm for consistent peak picking and FDR-controlled scoring. DreamDIAlignR outperformed state-of-the-art MBR methods, identifying up to 25.6% more quantitatively changing proteins on a benchmark dataset and 38.5% more on a cancer dataset. Additionally, DreamDIAlignR establishes an improved methodology for performing MBR compatible with existing DIA analysis tools, thereby enhancing the overall quality of DIA analysis.