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Exploration of peptides bound to MHC class I molecules in melanoma
Author(s) -
Pritchard Antonia L.,
Hastie Marcus L.,
Neller Michelle,
Gorman Jeffrey J.,
Schmidt Chris W.,
Hayward Nicholas K.
Publication year - 2015
Publication title -
pigment cell and melanoma research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.618
H-Index - 105
eISSN - 1755-148X
pISSN - 1755-1471
DOI - 10.1111/pcmr.12357
Subject(s) - peptide , human leukocyte antigen , epitope , mhc class i , chemistry , major histocompatibility complex , mass spectrometry , microbiology and biotechnology , computational biology , biology , biochemistry , antigen , chromatography , immunology , gene
Summary 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.