Schematic representation of a strategy to systematically discover tumor neoantigens. Tumor-specific mutations in cancer samples are detected using WES or WGS and identified through the application of mutation calling algorithms (such as Mutect).³³  Next, candidate neoepitopes can be predicted using well-validated algorithms (NetMHCpan), and their identification can be refined by experimental validation for peptide-HLA binding and by confirmation of gene expression at the RNA level. These candidate neoantigens can be subsequently tested for their ability to stimulate tumor-specific T-cell responses.