SPR was performed using the Biacore T200 instrument as previously described [22 (link)]. eEF2 protein was diluted in 10 mM sodium acetate (pH 4.5) at a concentration of 15 μg/mL for immobilization. After immobilizing about 4 µg eEF2 protein on a CM5 chip (Cat#BR-1005–30, GE Healthcare), TSN or DDD107498 (dissolved in PBS with 0.1% DMSO) were perfused through the ligand channels at different concentrations ranging from 10 nM to 1 µM. The T200 evaluation software was used to analyze the binding affinity between eEF2 with TSN or DDD107498.
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