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Immunopeptidomics is a field that studies all peptides presented by cells in the antigen presentation pathway. It mainly focuses on peptides on MHC-I or II proteins (such as HLA, the expression product of human MHC) on the cell surface. Identification of tumor antigens with immunopeptidomics, which adopts technologies such as immunoaffinity purification and high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS), is regarded as a tool for efficiently screening naturally presented peptides and neoepitopes. Through large-scale immunopeptidomics, it can reveal hotspots in the human proteome that have better recognition and binding capabilities for peptide epitopes, including the identification of neoepitopes derived from mutations. This is conducive to the treatment of gene mutation-derived tumors and the realization of individualized precision treatment.