Fig. 3. Validation and functional characterization of mKRAS-specific TCRs.

a FACS histogram plots demonstrating p-HLA multimer staining following lentiviral engineering of J^ASP90 reporter cells with TCRA3V, TCRA11V, and TCRB7R constructs (red) vs TCRαβ^null controls (black). b J^ASP90 reporter assay to evaluate antigen recognition. J^ASP90 reporter cells expressing TCRA3V, TCRA11V, or TCRB7R were cocultured with HLA-matched monoallelic K562 cell lines pulsed with titrated concentrations of WT KRAS or mKRAS peptides. TCR signaling is indicated by eGFP expression upon NFAT activation. TCR avidity was determined as the mean peptide concentration required to achieve 50% NFAT activation of TCR-engineered J^ASP90 reporter cells. Specific Activity (%) = (%GFP_Test – %GFP_Min) / (%GFP_Max – %GFP_Min) x 100. Representative experiments of three independent evaluations are shown. Source data are provided as a Source Data file.