|Anti-human PD-1 inhibits the binding of PD-L1. The graphs show two sets of data. First, recombinant human PD-L1-Fc binds to immobilized human PD-1-Fc that has been coated overnight on a 96-well plate (Cyan lines). PD-1-Fc was coated at 1 µg/mL, and increasing concentrations of PD-L1-Fc (cyan diamonds) or paired PBS wells (cyan circles) were incubated overnight. The bound recombinant PD-L1-Fc was detected using biotin anti-human PD-L1 and Avidin HRP, followed by TMB. After verifying that PD-L1-Fc binds to immobilized PD-1-Fc, a second experiment was performed in which the coated PD-1-Fc was pre-incubated with increasing concentrations of anti-human PD-1 (clone EH12.2H7, purple squares) or isotype control (clone MOPC-21, purple circles), followed by incubation with a fixed concentration of PD-L1 (1 µg/mL). Clone EH12.2H7 inhibits PD-1/PD-L1 interaction in a dose dependent manner.