3x104 A431 cells were injected subcutaneously into the right shoulder of 4-6-week-old BALB/c nude mice, and when the average tumor size of the mice grew to 140 ~ 150mm3, they were randomly divided into 5 cells in each group. At day 0, day 7, day 14, day 21, intravenous injection of a blank control (blank buffer solution) and a dose of 10mg/kg antibody drug conjugate was administered. The tumor volume was measured at day 0, day 7, day 14, day 21, day 24, day 28, day 32, day 36 and the tumor growth inhibition rate on day 28 of d was calculated.

N87 cells were used as in vitro pharmacodynamic detection systems. An appropriate number of tumor cell lines were inoculated in 96-well plates, incubated in carbon dioxide incubators for 24 hours, and then treated with medicine. The drug was diluted in a medium (the initial concentration of ADC drug was 500nM, the dilution factor was 7 times, there were 8 concentration points, the theoretical coupling ratio (DAR) of toxin and antibody was 8:1, and the actual coupling ratio was roughly 7.5:1, so the initial concentration of toxin was 4.0 uM, 7 times concentration gradient dilution, 8 concentration points). After mixing, it was added into the corresponding cell holes and incubated at 37°C in carbon dioxide incubator for 5 days. After 5 days, 20uL MTS was added to each well (Promega, G3581) reacted for 2 hours, and the absorption value reading at 490nm was taken with an enzyme label. By detecting the activity of dehydrogenase in mitochondria, IC50 was calculated to evaluate the inhibitory effect of ADC drugs on the proliferation of tumor cells.

SW620 cells were used as in vitro pharmacodynamic detection systems. An appropriate number of tumor cell lines were inoculated in 96-well plates, incubated in carbon dioxide incubators for 24 hours, and then treated with medicine. The drug was diluted in a medium (the initial concentration of ADC drug was 500nM, the dilution factor was 7 times, there were 8 concentration points, the theoretical coupling ratio (DAR) of toxin and antibody was 8:1, and the actual coupling ratio was roughly 7.5:1, so the initial concentration of toxin was 4.0 uM, 7 times concentration gradient dilution, 8 concentration points). After mixing, it was added into the corresponding cell holes and incubated at 37°C in carbon dioxide incubator for 5 days. After 5 days, 20uL MTS was added to each well (Promega, G3581) reacted for 2 hours, and the absorption value reading at 490nm was taken with an enzyme label. By detecting the activity of dehydrogenase in mitochondria, IC50 was calculated to evaluate the inhibitory effect of ADC drugs on the proliferation of tumor cells.

Fadu cells were used as in vitro pharmacodynamic detection systems. An appropriate number of tumor cell lines were inoculated in 96-well plates, incubated in carbon dioxide incubators for 24 hours, and then treated with medicine. The drug was diluted in a medium (the initial concentration of ADC drug was 500nM, the dilution factor was 7 times, there were 8 concentration points, the theoretical coupling ratio (DAR) of toxin and antibody was 8:1, and the actual coupling ratio was roughly 7.5:1, so the initial concentration of toxin was 4.0 uM, 7 times concentration gradient dilution, 8 concentration points). After mixing, it was added into the corresponding cell holes and incubated at 37°C in carbon dioxide incubator for 5 days. After 5 days, 20uL MTS was added to each well (Promega, G3581) reacted for 2 hours, and the absorption value reading at 490nm was taken with an enzyme label. By detecting the activity of dehydrogenase in mitochondria, IC50 was calculated to evaluate the inhibitory effect of ADC drugs on the proliferation of tumor cells.

BXPC-3 cells were used as in vitro pharmacodynamic detection systems. An appropriate number of tumor cell lines were inoculated in 96-well plates, incubated in carbon dioxide incubators for 24 hours, and then treated with medicine. The drug was diluted in a medium (the initial concentration of ADC drug was 500nM, the dilution factor was 7 times, there were 8 concentration points, the theoretical coupling ratio (DAR) of toxin and antibody was 8:1, and the actual coupling ratio was roughly 7.5:1, so the initial concentration of toxin was 4.0 uM, 7 times concentration gradient dilution, 8 concentration points). After mixing, it was added into the corresponding cell holes and incubated at 37°C in carbon dioxide incubator for 5 days. After 5 days, 20uL MTS was added to each well (Promega, G3581) reacted for 2 hours, and the absorption value reading at 490nm was taken with an enzyme label. By detecting the activity of dehydrogenase in mitochondria, IC50 was calculated to evaluate the inhibitory effect of ADC drugs on the proliferation of tumor cells.

A431 cells were used as in vitro pharmacodynamic detection systems. An appropriate number of tumor cell lines were inoculated in 96-well plates, incubated in carbon dioxide incubators for 24 hours, and then treated with medicine. The drug was diluted in a medium (the initial concentration of ADC drug was 500nM, the dilution factor was 7 times, there were 8 concentration points, the theoretical coupling ratio (DAR) of toxin and antibody was 8:1, and the actual coupling ratio was roughly 7.5:1, so the initial concentration of toxin was 4.0 uM, 7 times concentration gradient dilution, 8 concentration points). After mixing, it was added into the corresponding cell holes and incubated at 37°C in carbon dioxide incubator for 5 days. After 5 days, 20uL MTS was added to each well (Promega, G3581) reacted for 2 hours, and the absorption value reading at 490nm was taken with an enzyme label. By detecting the activity of dehydrogenase in mitochondria, IC50 was calculated to evaluate the inhibitory effect of ADC drugs on the proliferation of tumor cells.

SW620 cells were used as in vitro pharmacodynamic detection systems. An appropriate number of tumor cell lines were inoculated in 96-well plates, incubated in carbon dioxide incubators for 24 hours, and then treated with medicine. The drug was diluted in a medium (the initial concentration of ADC drug was 500nM, the dilution factor was 7 times, there were 8 concentration points, the theoretical coupling ratio (DAR) of toxin and antibody was 8:1, and the actual coupling ratio was roughly 7.5:1, so the initial concentration of toxin was 4.0 uM, 7 times concentration gradient dilution, 8 concentration points). After mixing, it was added into the corresponding cell holes and incubated at 37°C in carbon dioxide incubator for 5 days. After 5 days, 20uL MTS was added to each well (Promega, G3581) reacted for 2 hours, and the absorption value reading at 490nm was taken with an enzyme label. By detecting the activity of dehydrogenase in mitochondria, IC50 was calculated to evaluate the inhibitory effect of ADC drugs on the proliferation of tumor cells.

N87 cells were used as in vitro pharmacodynamic detection systems. An appropriate number of tumor cell lines were inoculated in 96-well plates, incubated in carbon dioxide incubators for 24 hours, and then treated with medicine. The drug was diluted in a medium (the initial concentration of ADC drug was 500nM, the dilution factor was 7 times, there were 8 concentration points, the theoretical coupling ratio (DAR) of toxin and antibody was 8:1, and the actual coupling ratio was roughly 7.5:1, so the initial concentration of toxin was 4.0 uM, 7 times concentration gradient dilution, 8 concentration points). After mixing, it was added into the corresponding cell holes and incubated at 37°C in carbon dioxide incubator for 5 days. After 5 days, 20uL MTS was added to each well (Promega, G3581) reacted for 2 hours, and the absorption value reading at 490nm was taken with an enzyme label. By detecting the activity of dehydrogenase in mitochondria, IC50 was calculated to evaluate the inhibitory effect of ADC drugs on the proliferation of tumor cells.

Fadu cells were used as in vitro pharmacodynamic detection systems. An appropriate number of tumor cell lines were inoculated in 96-well plates, incubated in carbon dioxide incubators for 24 hours, and then treated with medicine. The drug was diluted in a medium (the initial concentration of ADC drug was 500nM, the dilution factor was 7 times, there were 8 concentration points, the theoretical coupling ratio (DAR) of toxin and antibody was 8:1, and the actual coupling ratio was roughly 7.5:1, so the initial concentration of toxin was 4.0 uM, 7 times concentration gradient dilution, 8 concentration points). After mixing, it was added into the corresponding cell holes and incubated at 37°C in carbon dioxide incubator for 5 days. After 5 days, 20uL MTS was added to each well (Promega, G3581) reacted for 2 hours, and the absorption value reading at 490nm was taken with an enzyme label. By detecting the activity of dehydrogenase in mitochondria, IC50 was calculated to evaluate the inhibitory effect of ADC drugs on the proliferation of tumor cells.

BXPC-3 cells were used as in vitro pharmacodynamic detection systems. An appropriate number of tumor cell lines were inoculated in 96-well plates, incubated in carbon dioxide incubators for 24 hours, and then treated with medicine. The drug was diluted in a medium (the initial concentration of ADC drug was 500nM, the dilution factor was 7 times, there were 8 concentration points, the theoretical coupling ratio (DAR) of toxin and antibody was 8:1, and the actual coupling ratio was roughly 7.5:1, so the initial concentration of toxin was 4.0 uM, 7 times concentration gradient dilution, 8 concentration points). After mixing, it was added into the corresponding cell holes and incubated at 37°C in carbon dioxide incubator for 5 days. After 5 days, 20uL MTS was added to each well (Promega, G3581) reacted for 2 hours, and the absorption value reading at 490nm was taken with an enzyme label. By detecting the activity of dehydrogenase in mitochondria, IC50 was calculated to evaluate the inhibitory effect of ADC drugs on the proliferation of tumor cells.

A431 cells were used as in vitro pharmacodynamic detection systems. An appropriate number of tumor cell lines were inoculated in 96-well plates, incubated in carbon dioxide incubators for 24 hours, and then treated with medicine. The drug was diluted in a medium (the initial concentration of ADC drug was 500nM, the dilution factor was 7 times, there were 8 concentration points, the theoretical coupling ratio (DAR) of toxin and antibody was 8:1, and the actual coupling ratio was roughly 7.5:1, so the initial concentration of toxin was 4.0 uM, 7 times concentration gradient dilution, 8 concentration points). After mixing, it was added into the corresponding cell holes and incubated at 37°C in carbon dioxide incubator for 5 days. After 5 days, 20uL MTS was added to each well (Promega, G3581) reacted for 2 hours, and the absorption value reading at 490nm was taken with an enzyme label. By detecting the activity of dehydrogenase in mitochondria, IC50 was calculated to evaluate the inhibitory effect of ADC drugs on the proliferation of tumor cells.