Monitoring the binding of hIL-6 or vIL-6 to immobilized sgp130 using the biosensor system BIAcore 2000.
The sgp130 was immobilized at a concentration of 18.2 ng/mm2 on the CM5 biosensor chip. (A) hIL-6 (50 μg/mL) alone or hIL-6 (50 μg/mL) plus sIL-6R (20 μg/mL), (B) MBPvIL-6 (50 μg/mL) alone or MBPvIL-6 (50 μg/mL) plus sIL-6R (20 μg/mL) was passed over the sensor surface at a flow rate of 10 μL/min in PBS. Reagents were incubated for 1 hour prior to assay. (C) Overlay of sensorgrams showing kinetic analysis of vIL-6 with sgp130. The sgp130 was immobilized at a concentration of 1.5 ng/mm2. MBPvIL-6 (100, 200, 400, and 800 μg/mL) was passed over the sensor surface. Control MBP (800 μg/mL) did not show detectable affinity for sgp130. Representative data of 4 independent experiments are shown.
