A simple strategy for one-step construction of bienzyme biosensor by in-situ formation of biocomposite film through electrodeposition.
One-step construction of horseradish peroxidase (HRP) and glucose oxidase (GOD) bienzyme biosensor was established based on a simple and controllable electrodeposition approach by in-situ formation of biocomposite film on electrode. With hydrogen peroxide (H(2)O(2)) instantly generated by GOD-catalyzed oxidation of glucose, quantitative measurement of glucose could be achieved. In the proposed electrodeposition strategy, Concanavalin A (Con A) was applied as bifunctional cross-linker for in-situ incorporating of HRP-GOD in biocompatible chitosan (CS) matrix due to its biospecific affinity interaction with CS and glycoproteins. Scanning electron microscopy showed that the HRP-GOD/Con A/CS biocomposite film possessed highly porous surface. The developed bienzyme biosensor exhibited a fast amperometric response for the determination of glucose. The linear response of the developed biosensor for the determination of glucose ranged from 1.0x10(-6) to 2.2x10(-4) M with a detection limit of 6.7x10(-7) M. The biosensor can be used to determine glucose in serum directly. The biosensor presented high stability owing to the design of the introduction of Con A and instant generation of H(2)O(2) with bienzyme system. The one-step construction of biosensor based on non-manual technique in biocompatible environment was direct and facile.
Li
Feng
F
Key Laboratory of Eco-Chemical Engineering, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, PR China.
Wang
Zhen
Z
Chen
Wei
W
Zhang
Shusheng
S
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods* / Q000706:statistics & numerical data; D001786:Blood Glucose / Q000032:analysis; D048271:Chitosan; D020099:Coated Materials, Biocompatible; D055664:Electrochemical Techniques; D004800:Enzymes, Immobilized; D005949:Glucose Oxidase; D006735:Horseradish Peroxidase; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008855:Microscopy, Electron, Scanning; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D001786:Blood Glucose; D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D048271:Chitosan; D005949:Glucose Oxidase; D006735:Horseradish Peroxidase