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An optical biosensor for kinetic analysis of soluble Interleukin-1 receptor I binding to immobilized Interleukin-1alpha
Authors:Wu Bao-Yan  Wang Yan-Yan  Li Jing  Song Zhao  Huang Jia-Dong  Wang Xin-Sheng  Chen Qiang
Institution:College of Life Sciences, Nankai University, Tianjin 300071, China
Abstract:We report here the development of an optical biosensor based on the resonant mirror for kinetic analysis of soluble Interleukin-1 receptor I (sIL-1R I) in solution binding to immobilized Interleukin-1α (IL-1α). IL-1α was immobilized through its surface amine groups via amide bonds with the carboxyl groups of the carboxymethyl dextran (CMD) on cuvette surface. The interaction of sIL-1R I and IL-1α was monitored in real time. Evaluation of the binding curves allowed the analysis of the binding kinetics. The linear range of sIL-1R I in solution was over a range of 100-1600 nM (R = 0.9962). Equilibrium dissociation constant (KD) was derived by Scatchard plot analysis for sIL-1R I binding to immobilized IL-1α. For this assay, the KD was 2.6 × 10−6 M. The CMD cuvette modified by IL-1α was successfully regenerated using 10 mM HCl, and the same sensing surface was used repeatedly for the interaction analysis.
Keywords:Optical biosensor  Kinetic analysis  Soluble Interleukin-1 receptor I  Interleukin-1α
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