Abstract
Male infertility is fundamentally rooted in developmental defects of germ cells and associated molecular dysregulation, yet the underlying mechanisms remain poorly understood. Data-independent acquisition (DIA) has emerged as a powerful tool in discovery proteomics, enabling the identification of disease biomarkers, therapeutic targets, and molecular pathways with high precision and reproducibility. To analyse the aberrant regulatory events in human sperm proteins associated with male reproductive disorders in a high-throughput, reproducible, and reliable manner, we employed the Orbitrap Astral mass spectrometer, which independently operates Orbitrap Full Scan and Astral MS/MS to generate high-resolution full-scan spectra and high-quality secondary maps. Leveraging the principles of DIA technology, we constructed the most comprehensive human sperm proteomic expression profile reported to date, encompassing 9,309 proteins, 198,153 unique precursors, 154,062 modified peptides, and 145,355 peptides. Moreover, this data is derived from two types of sperm samples to facilitate the exploration of disease-specific proteins. Our data serve as a valuable resource for analysing spermatozoa protein content, facilitating deeper insights into male reproduction.