| Assay Method Information | |
| | Measurement of Molecular Interactions Using Surface Plasmon Resonance (SPR) |
| Description: | The instrument used was the Biacore T200 (manufactured by Cytiva). The CRBN truncate (318-426) was obtained by expressing a construct with an Avi-tag added at the C-terminus. The sensor chip used was Sensor Chip SA (manufactured by Cytiva). The CRBN truncate was immobilized using the affinity between SA (streptavidin) on the sensor chip and the Avi-tag. The running buffer was a mixture of 10 mM HEPES pH 7.5, 150 mM NaCl, 1 mM TCEP, 0.005% Tween 20, and 2% DMSO. Under 25° C. conditions, solutions of various concentrations of test compounds (compounds of the examples above) were flowed at 30 L/min over the sensor chip with the immobilized CRBN truncate, and the mass changes on the sensor chip were observed. Contact time and dissociation time were set at 60 seconds and 120 seconds, respectively. The observed data were analyzed using Biacore T200 Evaluation Software Version 2.0 (manufactured by Cytiva), and the Kd values of the compounds for the CRBN truncate were calculated. In this test, the upper limit for the Kd value measurement, based on the lower quantification limit, was 200 μM. |
| Affinity data for this assay | |
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