The proteins of bacteriophages were subjected to extraction and purification, target protein digestion, peptide separation, mass spectrometry detection, and data analysis. Firstly, a comprehensive and high-quality spectral library of peptides was constructed. DIA data acquisition was conducted based on the spectral library. During the acquisition process, all peptides within the window underwent secondary fragmentation, and the corresponding high-resolution secondary spectra were collected. The obtained data were analyzed by bioinformatics.