Peptides that bind MHC Class I are readily identified with a programmable, high-throughput method
A lentiviral pHAGE vector with a cytomegalovirus promoter plus an EF1α promoter driving EGFP–P2A-Purowas used as the backbone. The vector was digested with PstI and AgeI to excise the EF1α promoter, and the Gibson assembly method was used to insert a gBlock encoding a codon-optimized mouse mammary tumor virus gp70 signal peptide, filler region flanked by BsmBI sites and an IRES element.
Evaluating all top ESP predictions on the proteome. Of all ESPv1 proteome predictions, those with peptides with a binding probability of <0.05 or <0.1 were chosen: 84,580 peptides for HLA-A*02, HLA-B*08 and HLA-B*57 and 92,000 for HLA*03.
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