FIG. 3.

Cytotoxicity assay. (A) Cytotoxicity assay by cell viability assessment was conducted using CCK-8 We prepared 96-well microplates by plating CHO/dPDPN cells at 6000 cells/200 μL/well. The plates were incubated for 24 hours at 37°C in the presence of 5% CO₂, then serially diluted antibodies were added in a volume of 20 μL and cultured for 24 hours. After that, the culture medium was replaced with 100 μL of fresh medium with 10 μL CCK-8 solution. After a further 5-hour incubation in a CO₂ incubator, the absorbance at 450 nm was measured. The values are presented as mean ± SEM. Asterisks indicate statistical significance (**p < 0.01, Tukey–Kramer test). (B) Cytotoxicity assay by cell viability assessment was conducted using CCK-8 We prepared 96-well microplates by plating CHO/dPDPN and CHO-K1 cells at 6000 cells/200 μL/well. The plates were incubated for 24 hours at 37°C in the presence of 5% CO₂, then serially diluted antibodies were added in a volume of 20 μL and cultured for 72 hours. After that, the culture medium was replaced with 100 μL of fresh medium with 10 μL CCK-8 solution. After a further 6-hour incubation in a CO₂ incubator, the absorbance at 450 nm was measured. Values are presented as mean ± SEM. Asterisks indicate statistical significance (**p < 0.01, Tukey–Kramer test). n.s., not significant; CCK-8, Cell Cloning Kit-8; SEM, standard error of mean.