Cytotoxic effects of aTCRA6 DAR2 and aTCRA6 DAR52-8 ADCs were estimated by exposing Jurkat WT and Jurkat TCRA6 T cells to different ADC concentrations. Cell viability was analyzed 72 h post ADC addition by a fluorometric method using CellTiter-Blue Cell Viability Assay (Promega, Wisconsin, USA) according to the manufacturer's instructions. Briefly, cells were seeded in a 96-well plate (1 × 104 cells/well) with the desired ADC concentrations ranging from 0.06-400 nM (for DAR2 ADC) or 0.005-30 nM (for DAR52-8 ADC) in a serial dilution. After 72 h, redox dye (resazurin) was added to the cells, and the plate was incubated for 3-4 h. Fluorescence of resorufin (560Ex/590Em) was recorded using CLARIOstar plus microplate reader (BMG LABTECH, Offenburg, Germany). Cell proliferation was normalized to untreated control cell fluorescence values. The resulting curves were fitted with a variable slope four-parameter fit, and EC50s were calculated using GraphPad Prism.

Cytotoxic effects of aTCRA6 DAR2 and aTCRA6 DAR52-8 ADCs were estimated by exposing Jurkat WT and Jurkat TCRA6 T cells to different ADC concentrations. Cell viability was analyzed 72 h post ADC addition by a fluorometric method using CellTiter-Blue Cell Viability Assay (Promega, Wisconsin, USA) according to the manufacturer's instructions. Briefly, cells were seeded in a 96-well plate (1 × 104 cells/well) with the desired ADC concentrations ranging from 0.06-400 nM (for DAR2 ADC) or 0.005-30 nM (for DAR52-8 ADC) in a serial dilution. After 72 h, redox dye (resazurin) was added to the cells, and the plate was incubated for 3-4 h. Fluorescence of resorufin (560Ex/590Em) was recorded using CLARIOstar plus microplate reader (BMG LABTECH, Offenburg, Germany). Cell proliferation was normalized to untreated control cell fluorescence values. The resulting curves were fitted with a variable slope four-parameter fit, and EC50s were calculated using GraphPad Prism.

Cytotoxic effects of aTCRA6 DAR2 and aTCRA6 DAR52-8 ADCs were estimated by exposing Jurkat WT and Jurkat TCRA6 T cells to different ADC concentrations. Cell viability was analyzed 72 h post ADC addition by a fluorometric method using CellTiter-Blue Cell Viability Assay (Promega, Wisconsin, USA) according to the manufacturer's instructions. Briefly, cells were seeded in a 96-well plate (1 × 104 cells/well) with the desired ADC concentrations ranging from 0.06-400 nM (for DAR2 ADC) or 0.005-30 nM (for DAR52-8 ADC) in a serial dilution. After 72 h, redox dye (resazurin) was added to the cells, and the plate was incubated for 3-4 h. Fluorescence of resorufin (560Ex/590Em) was recorded using CLARIOstar plus microplate reader (BMG LABTECH, Offenburg, Germany). Cell proliferation was normalized to untreated control cell fluorescence values. The resulting curves were fitted with a variable slope four-parameter fit, and EC50s were calculated using GraphPad Prism.

Cytotoxic effects of aTCRA6 DAR2 and aTCRA6 DAR52-8 ADCs were estimated by exposing Jurkat WT and Jurkat TCRA6 T cells to different ADC concentrations. Cell viability was analyzed 72 h post ADC addition by a fluorometric method using CellTiter-Blue Cell Viability Assay (Promega, Wisconsin, USA) according to the manufacturer's instructions. Briefly, cells were seeded in a 96-well plate (1 × 104 cells/well) with the desired ADC concentrations ranging from 0.06-400 nM (for DAR2 ADC) or 0.005-30 nM (for DAR52-8 ADC) in a serial dilution. After 72 h, redox dye (resazurin) was added to the cells, and the plate was incubated for 3-4 h. Fluorescence of resorufin (560Ex/590Em) was recorded using CLARIOstar plus microplate reader (BMG LABTECH, Offenburg, Germany). Cell proliferation was normalized to untreated control cell fluorescence values. The resulting curves were fitted with a variable slope four-parameter fit, and EC50s were calculated using GraphPad Prism.