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Presentation of tumor-specific or tumor-associated peptides by HLA class I molecules to CD8 +  T cells is the foundation of epitope-centric cancer immunotherapies. While often  in silico  HLA binding predictions or  in vitro  immunogenicity assays are utilized to select candidates, mass spectrometry-based immunopeptidomics is currently the only method providing a direct proof of actual cell surface presentation. Despite much progress in the last decade, identification of such HLA-presented peptides remains challenging. Here we review typical workflows and current developments in the field of immunopeptidomics, highlight the challenges which remain to be solved and emphasize the importance of direct target validation for clinical immunotherapy development.

The Importance of Being Presented: Target Validation by Immunopeptidomics for Epitope-Specific Immunotherapies