Fig. 5 | Nature Communications

Fig. 5

From: Chemically triggered drug release from an antibody-drug conjugate leads to potent antitumour activity in mice

Fig. 5

Tetrazine-triggered drug release from tc-ADC in vitro. a HPLC-QTOF-MS analysis of tc-ADC showing, after deconvolution, the fully MMAE-conjugated monomer (30,595.50 Da) and, after a 1 h reaction with 3 in PBS at 37 °C, the monomer with two pyridazine derivatives and no MMAE (31719.60 Da: 80%) and the monomer with two pyridazine derivatives and one MMAE still attached (32480.80 Da; 20%). b HPLC-QTOF-MS analysis of deproteinized mixtures of tc-ADC and 3 (in triplicate) after 10 min (orange), 1 h (blue) and 20 h (magenta) incubation in 50% serum at 37 °C; c quantification of recovered MMAE using calibration curves. Cytotoxicity of combined tc-ADC and 3 in d LS174T cells (EC50 = 185 pM) and e OVCAR-3 cells (EC50 = 35 pM) in vitro, in comparison with only tc-ADC or 3; free MMAE (EC50 = 277 and 39 pM) was used as positive control. As the ADC activation was not performed on the cell surface but in solution in the presence of the cells, similar results were obtained for the TAG72-negative HT-29 cells, see Supplementary Table 1 and Supplementary Fig. 12

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