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Indeed, it has been recently proven the relevance of the position of the mutated amino acid in neoAg choice . HLA class I molecules expressed by patients had been recognized using sequencing knowledge and all potential peptides containing mutated residues were chosen for prediction of peptide binding to HLA. By using a threshold of 500 nM in affinity and a % Rank Table 1, Supplementary Table S4). NeoAg-specific responses had been evaluated at day 21 of co-culture by using a human IFN-γ ELISPOT set (BD-Biosciences) following manufacturer’s instructions. Expanded CD8 T cells (5 x 104/well) were stimulated with peptides (10 µg/mL) and DC (104/well) for 24 h.
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Prediction of binding to HLA class I molecules of mutated and wild type peptides. Different criteria have been used to choose out predicted peptides.
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