| Assay Method Information | |
| | Surface Plasmon Resonance (SPR) |
| Description: | Binding kinetics were determined by surface plasmon resonance (SPR) using a Biacore S51 (Biacore, Uppsala, Sweden). MPO (50 μg/ml dissolved in 10 mM sodium acetate, pH 5.0) was immobilized onto the surface of CM5 sensor chips (Biacore) using surface amine coupling. One of the spots on the sensor surface was left without MPO to control for nonspecific binding. The signal observed in response to analyte binding was as expected linearly related to the amount of immobilized ligand, and 10,000 response units was routinely used to characterize compoundbinding (data not shown). Compounds were dissolved in binding buffer (10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM Na2EDTA, 0.005% (w/v) surfactant P20, 1% (v/v) DMSO final), and association was assessed during 60-210-s injections. After this time, analyte injection was terminated, and the chip surface was perfused with binding buffer at 30 μl/min for 4-12 min to monitor compound dissociation. |
| Affinity data for this assay | |
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