| Assay Method Information | |
| | PD1-PDL1 HTRF Binding Activity Test |
| Description: | The effect of compounds of the examples of the present invention on the PD-1/PD-L1 interaction was determined by the PD-1/PD-L1 binding assay kit from Cisbio (#64ICP01PEG or 64ICP01PEH). The detailed experimental process was as follows:1) pre-diluted compound solution, 4 μL of Tag1-PD-L1 and 4 μL of Tag2-PD1 were each added to a 384-well plate;2) after the mixture was incubated at room temperature for 15 min, 5 μL of anti-Tag1-Eu3+ antibody and 5 μL of anti-Tag2-XL665 antibody were then added;3) after being incubated for 2 hrs at room temperature or overnight at 4° C., the plate was read on Envision of Perkin Elmer; readings at 665 nm and 620 nm were recorded, and the ratio of the two readings was taken as the reading of each well;4) the reading of each well after compound treatment was compared with the reading of DMSO treated wells to obtain the percent inhibition of the compound; and5) IC50 values of compounds of the examples of the present invention were determined by non-linear regression analysis of percent inhibition at different compound concentrations. |
| Affinity data for this assay | |
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