Exploration of peptides bound to MHC class I molecules in melanoma. (5th March 2015)

• Record Type: Journal Article
• Authors: Pritchard, Antonia L.; Hastie, Marcus L.; Neller, Michelle; Gorman, Jeffrey J.; Schmidt, Chris W.; Hayward, Nicholas K.
• Is Part Of: Pigment cell & melanoma research. Volume 28:Number 3(2015:May)
• Page Start: 281
• Page End: 294
• Publication Date: 2015-03-05
• Subjects: Melanoma -- Periodicals; Chromatophores -- Periodicals; Animal pigments -- Periodicals; 616.99477
• DOI: 10.1111/pcmr.12357
• Languages: English
• ISSNs: 1755-1471

Abstract

<abstract abstract-type="main" id="pcmr12357-abs-0001"> <title>Summary</title> <p>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; mRNA 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 <italic>ex vivo</italic> by one patient's T cells.</p> </abstract>