Fig. 6
Antibody–drug conjugate P1C1TM-PNU mediates cytotoxicity of tumour cells presenting p53125–134/A24 pMHC. a P1C1TM and an isotype control human IgG1 was conjugated with the pH-dependent dye, pHrodo Red and incubated with HT29 cells at 37 °C or on ice. At different time points, cells were washed with cold PBS and internalization of pHrodo Red-labelled antibodies was assessed by flow cytometry. Data are representative of two independent experiments. b HT29 cells were incubated with varying concentrations of P1C1TM in the presence or absence of four different anti-human IgG secondary antibody–drug conjugates in equimolar ratios. Cell viability was assessed after 3 days by an MTS assay. Data are means of triplicates ± SEM. c Cytotoxic drug PNU-159682 was directly conjugated to P1C1TM (P1C1TM-PNU) and assessed for binding to HT29 and HepG2 cells by flow. d HLA-A24 expressing cell lines expressing mutant p53 (HT29, MDA-MB-231-A24) and wild-type p53 (HepG2, A549-A24, MCF7-A24) were incubated with varying concentrations of P1C1TM only or the P1C1TM-PNU antibody–drug conjugate. Cell viability was assessed by MTS assay after 3 days. Data are means of triplicates ± SEM.
