| Assay Method Information | |
| | PHGDH SPR Assay |
| Description: | The binding affinities of compounds towards PHGDH were assayed using the SPR-based Biacore T200 instrument (GE Healthcare). PHGDH was immobilized on a CM5 sensor chip by using standard amine-coupling at 25℃ with running buffer PBS-P (20 mM phosphate buffer, 2.7 mM NaCl, 137 mM KCl, 0.05% surfactant P-20, pH 7.4), respectively, as described previously. A reference flow cell was activated and blocked in the absence of PHGDH. In the direct binding experiments between PHGDH and compounds, PHGDH immobilization level was fixed at 800 response units (RU), and then different concentrations of compounds containing 5% DMSO were serially injected into the channel to evaluate binding affinity. Regeneration was achieved by extended washing with the running buffer after each sample injection. |
| Affinity data for this assay | |
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