A total of 625 cells (WISH) in a volume of 25 uL were seeded in a 384-well flat bottom white polystyrene plate the day before the actual assay started. ADC and free drugs were formulated at 2x starting concentration in cell culture medium and filtered through 0.22um cellulose acetate filtered 2 ml centrifuge tubes. Filter sterilized samples were serial diluted (1:3) under sterile conditions and 25 uL of each dilution was added onto cells in triplicates. Plates were cultured at 37 °C in a C02 incubator for 120 hours. For cell viability measurement, 30 uL of Cell Titer-Glo@ reagent (PromegaT M Corp, Madison, WI) was added into each well, and plates processed as per product instructions. Relative luminescence was measured.

A total of 625 cells (MDA-MB-468) in a volume of 25 uL were seeded in a 384-well flat bottom white polystyrene plate the day before the actual assay started. ADC and free drugs were formulated at 2x starting concentration in cell culture medium and filtered through 0.22um cellulose acetate filtered 2 ml centrifuge tubes. Filter sterilized samples were serial diluted (1:3) under sterile conditions and 25 uL of each dilution was added onto cells in triplicates. Plates were cultured at 37 °C in a C02 incubator for 120 hours. For cell viability measurement, 30 uL of Cell Titer-Glo@ reagent (PromegaT M Corp, Madison, WI) was added into each well, and plates processed as per product instructions. Relative luminescence was measured.

A total of 625 cells (OVCAR-3) in a volume of 25 uL were seeded in a 384-well flat bottom white polystyrene plate the day before the actual assay started. ADC and free drugs were formulated at 2x starting concentration in cell culture medium and filtered through 0.22um cellulose acetate filtered 2 ml centrifuge tubes. Filter sterilized samples were serial diluted (1:3) under sterile conditions and 25 uL of each dilution was added onto cells in triplicates. Plates were cultured at 37 °C in a C02 incubator for 120 hours. For cell viability measurement, 30 uL of Cell Titer-Glo@ reagent (PromegaT M Corp, Madison, WI) was added into each well, and plates processed as per product instructions. Relative luminescence was measured.

A total of 625 cells (NCI-H292) were plated in 90 ul cell culture medium per well of a 96-well black/clear flat bottom TC-treated imaging microplate and cultured overnight. A ten-fold starting concentration of ADCs, a respective serial dilution (1:4) were prepared in cell culture medium briefly before use. Wells were supplied with 10 ul ADC or compound solution. Treatment was performed in technical triplicates. The plates were subsequently incubated for 144h. For subsequent cell viability measurement, 100 ul Cell Titer-Glo reagent was pipetted in each well, and plates were further processed according the manufacturer's instructions.

A total of 625 cells (NCI-H292) were plated in 90 ul cell culture medium per well of a 96-well black/clear flat bottom TC-treated imaging microplate and cultured overnight. A ten-fold starting concentration of ADCs, a respective serial dilution (1:4) were prepared in cell culture medium briefly before use. Wells were supplied with 10 ul ADC or compound solution. Treatment was performed in technical triplicates. The plates were subsequently incubated for 144h. For subsequent cell viability measurement, 100 ul Cell Titer-Glo reagent was pipetted in each well, and plates were further processed according the manufacturer's instructions.

A total of 625 cells (NCI-H292) were plated in 90 ul cell culture medium per well of a 96-well black/clear flat bottom TC-treated imaging microplate and cultured overnight. A ten-fold starting concentration of ADCs, a respective serial dilution (1:4) were prepared in cell culture medium briefly before use. Wells were supplied with 10 ul ADC or compound solution. Treatment was performed in technical triplicates. The plates were subsequently incubated for 144h. For subsequent cell viability measurement, 100 ul Cell Titer-Glo reagent was pipetted in each well, and plates were further processed according the manufacturer's instructions.