1. Exploration of peptides bound to MHC class I molecules in melanoma.
- Author
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Pritchard, Antonia L., Hastie, Marcus L., Neller, Michelle, Gorman, Jeffrey J., Schmidt, Chris W., and Hayward, Nicholas K.
- Subjects
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PEPTIDES , *MAJOR histocompatibility complex , *MELANOMA , *HIGH performance liquid chromatography , *MASS spectrometry , *EPITOPES , *T cells - Abstract
Advancements in high-resolution HPLC and mass spectrometry have reinvigorated the application of this technology to identify peptides eluted from immunopurified MHC class I molecules. Three melanoma cell lines were assessed using w6/32 isolation, peptide elution and HPLC purification; peptides were identified by mass spectrometry. A total of 13 829 peptides were identified; 83-87% of these were 8-11 mers. Only approximately 15% have been described before. Subcellular locations of the source proteins showed even sampling; m RNA expression and total protein length were predictive of the number of peptides detected from a single protein. HLA-type binding prediction for 10 078 9/10 mer peptides assigned 88-95% to a patient-specific HLA subtype, revealing a disparity in strength of predicted binding. HLA-B*27-specific isolation successfully identified some peptides not found using w6/32. Sixty peptides were selected for immune screening, based on source protein and predicted HLA binding; no new peptides recognized by antimelanoma T cells were discovered. Additionally, mass spectrometry was unable to identify several epitopes targeted ex vivo by one patient's T cells. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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