In each ultra-low attachment U bottom 384-well transparent plate patient-derived lung cancer cells AMB-LC-0003T were dispensed in triplicate at 500 cells/well in 40 p1l of M10018 (Aimedbio) media. After centrifugation at 250 g for 2 minutes, reaction was carried out at 37°C and 5% Co 2 for 24 hours. Thereafter, each antibody-drug conjugate was serially diluted 3-fold from 500 nM to 0.314 pM and the cells were treated therewith. The plate was reacted again at 37°C and 5% Co2 for 6 days. After a total of 7 days of reaction, spheroids were formed by the cells in the 384-well plate, so images of spheroids in each well were captured using Operetta CLS (PerkinElmer) according to a high-content screening method. To maintain the spheroid form of the cells in each well, washing was not performed before analysis. The captured images were converted into volume through additional analysis, and the spheroid volume values by concentration were standardized based on the spheroid treated with the lowest concentration of drug. Graphs were created with a log (inhibitor) vs. normalized response - Variable slope using GraphPad Prism 9.3.1, and IC5 o values were obtained.

In each ultra-low attachment U bottom 384-well transparent plate patient-derived lung cancer cells AMB-LC-0003T were dispensed in triplicate at 500 cells/well in 40 p1l of M10018 (Aimedbio) media. After centrifugation at 250 g for 2 minutes, reaction was carried out at 37°C and 5% Co 2 for 24 hours. Thereafter, each antibody-drug conjugate was serially diluted 3-fold from 500 nM to 0.314 pM and the cells were treated therewith. The plate was reacted again at 37°C and 5% Co2 for 6 days. After a total of 7 days of reaction, spheroids were formed by the cells in the 384-well plate, so images of spheroids in each well were captured using Operetta CLS (PerkinElmer) according to a high-content screening method. To maintain the spheroid form of the cells in each well, washing was not performed before analysis. The captured images were converted into volume through additional analysis, and the spheroid volume values by concentration were standardized based on the spheroid treated with the lowest concentration of drug. Graphs were created with a log (inhibitor) vs. normalized response - Variable slope using GraphPad Prism 9.3.1, and IC5 o values were obtained.