期刊
PIGMENT CELL & MELANOMA RESEARCH
卷 28, 期 3, 页码 -出版社
WILEY-BLACKWELL
DOI: 10.1111/pcmr.12357
关键词
melanoma; mass spectrometry; MHC class I; proteome; antigen; epitope; peptide
资金
- Cure Cancer Australia
- Rio Tinto Ride to Conquer Cancer
- NHMRC
- Bioplatforms Australia
- Queensland State Government through the Australian Government National Collaborative Infrastructure Scheme (NCRIS)
- Education Investment Fund (EIF)
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 13829 peptides were identified; 83-87% of these were 8-11mers. Only approximately 15% have been described before. Subcellular locations of the source proteins showed even sampling; mRNA expression and total protein length were predictive of the number of peptides detected from a single protein. HLA-type binding prediction for 10078 9/10mer 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.
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