High‐sensitivity HLA class I peptidome analysis enables a precise definition of peptide motifs and the identification of peptides from cell lines and patients’ sera

• DOI: 10.1002/pmic.201500445
• Publication Date: 2016-03-19T01:01:56Z
• Authors: Danilo Ritz; Andreas Gloger; Benjamin Weide; Claus Garbe; Dario Neri; Tim Fugmann
• Keywords: Identification

The characterization of peptides bound to human leukocyte antigen (HLA) class I is fundamental importance for understanding CD8+ T cell-driven immunological processes and the development immunomodulatory therapeutic strategies. However, until now, mass spectrometric analysis HLA-bound has typically required billions cells, still resulting in relatively few high-confidence peptide identifications. Capitalizing on recent developments spectrometry bioinformatics, we have implemented a methodology efficient recovery acid-eluted HLA after purification with pan-reactive antibody W6/32 identified total 27 862 unique high confidence (1% false discovery rate) from five cancer cell lines. More than 93% were eight 11 amino acids length contained signatures that excellent agreement published binding motifs. Furthermore, by purifying soluble complexes (sHLA) sera melanoma patients, up 972 could be identified, including melanoma-associated antigens already described literature. Knowledge peptidome should facilitate multiplex tetramer technology-based allow patient selection, stratification