Electrochemical surface plasmon resonance detection of enzymatic reaction in bilayer lipid membranes.
In this paper, electrochemical surface plasmon resonance (SPR) method was first used to detect enzymatic reaction in bilayer lipid membrane (BLM) based on immobilizing horseradish peroxidase (HRP) in the BLMs supported by the redox polyaniline (PAn) film. By SPR kinetic curve in situ monitoring the redox transformation of PAn film resulted from the reaction between HRP and PAn, the enzymatic reaction of HRP with H(2)O(2) was successfully analyzed by electrochemical SPR spectroscopy. The results show that this BLM supported on PAn film cannot only preserve the bioactivity of HRP immobilized in the membrane, but also provide a channel for the transfer of electrons between HRP and PAn on electrode surface. These characteristics enabled the development of SPR biosensor for sensitively detecting H(2)O(2). H(2)O(2) has been detected by electrochemical SPR spectroscopy in the concentration range of 5 x 10(-5)M to 2 x 10(-3)M. After each of detections, the SPR sensor surface was completely regenerated by electrochemically reducing the oxidized PAn to its reduced state. This method provides a novel route for enhancing the detection of small ligand of enzymatic reaction in BLM by electrochemical SPR spectroscopy.
Wang
Jianlong
J
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Changchun Jilin 130022, Graduate School of the Chinese Academy of Sciences, Beijing 100039, China.
Wang
Fuan
F
Chen
Hongjun
H
Liu
Xiaohua
X
Dong
Shaojun
S
D004563:Electrochemistry; D004800:Enzymes, Immobilized; D006046:Gold / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force; D008954:Models, Biological; D020349:Surface Plasmon Resonance; D013499:Surface Properties
D004800:Enzymes, Immobilized; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D006046:Gold; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase