| Assay Method Information | |
| | hPD-1/hPD-L1 binding assay |
| Description: | A negative control, a positive control and drug administration groups were set up, with two duplicate wells in each group. For the positive control group, added 2 μL diluent to a 96-well plate; 4 μL of PD-L1 and 4 uL of PD-1 as diluted according to the instructions; for the negative control group, added 6 μL diluent and 4 μL PD-L1 to a 96-well plate; for the administration group, 2 μL of the compound of the invention (or the positive compound BMS-202), 4 μL of PD-L1 and 4 μL of PD-1 were successively added to a 96-well plate. Sealed the plate with a sealing film, centrifuged at 1000 rpm for 1 minute, and incubated at room temperature for 15 minutes. Then mixed equal volumes of Anti-Tag-Eu3+ and Anti-tag-XL665 as diluted in buffer evenly, then added 10 μL of the mixture to each well, sealed the plate, centrifuged at 1000 rpm for 1 minute, and incubated at room temperature for 2 hours. Removed the sealing film, used EnVision to read the fluorescence intensity at 665 nm and 615 nm, and calculate ratio=Signal 665 nm/Signal 620 nm×104. IC50 of the compounds was calculated using Graphpad. In this experiment, BMS-202 in the patent WO2015034820 of BMS Company was selected as the positive drug. |
| Affinity data for this assay | |
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