Surface Plasmon Resonance

Kinetics of IgG1-sFcyRIIIa interaction were measured using a BIAcore 2000 instrument and CM5 sensor chips (BIACORE, Uppsala, Sweden). Anti-Tetra His antibodies (QIAGEN) were immobilized onto the chip using amine coupling kit (BIACORE) following the manufacturer's instructions. sFcyRIIIa was captured by the immobilized anti-His antibodies by injecting sFcyRIIIa at a flow rate of 5 ^l/min. IgG1 was diluted in HBS-EP buffer (0.01 M HEPES, 0.15 M NaCl, 3 mM EDTA, 0.005% Surfactant P20, pH 7.4) at six different concentrations (from 4.17 nM to 133.3 nM) and each diluted IgG1 was injected over the receptor-captured sensor surface at a flow rate of 5 ^.l/min. sFcyRIIIa and IgG1 bound to the sensor surface were removed by injecting 7.5 mM HCl at a flow rate 10 ^.l/min for 30 sec. The experiments were performed at 25°C with HBS-EP as running buffer. Buffer solution without IgG1 was injected over the receptor-captured sensor surface as a blank control. The data obtained by the injection of IgG1 were corrected for the blank control prior to data analysis.

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