One-step immobilization of glucose oxidase in a silica matrix on a Pt electrode by an electrochemically induced sol-gel process.
We demonstrate here that the electrochemical generation of hydroxyl ions and hydrogen bubbles can be used to induce the synthesis of enzyme- or protein-encapsulated 3D porous silica structure on the surface of noble metal electrodes. In the present work, the one-step synthesis of a glucose oxidase (GOD)-encapsulated silica matrix on a platinum electrode is presented. In this process, glucose oxidase was mixed with ethanol and TEOS to form a doped precursory sol solution. The electrochemically generated hydrogen bubbles at negative potentials assisted the formation of the porous structure of a GOD-encapsulated silica gel, and then the one-step immobilization of enzyme into the silica matrix was achieved. Scanning electron microscopy (SEM) and scanning electrochemical microscopy (SECM) characterizations showed that the GOD-encapsulated silica matrix adhered to the electrode surface effectively and had an interconnected porous structure. Because the pores started at the electrode surface, their sizes increased gradually along the distance away from the electrode and reached maximum at the solution side, and effective mass transport to the electrode surface could be achieved. The entrapped enzyme in the silica matrix retained its activity. The present glucose biosensor had a short response time of 2 s and showed a linear response to glucose from 0 to 10 mM with a correlation coefficient of 0.9932. The detection limit was estimated to be 0.01 mM at a signal-to-noise ratio of 3. The apparent Michaelis-Menten constant (K m app) and the maximum current density were determined to be 20.3 mM and 112.4 microA cm-2, respectively. The present method offers a facile way to fabricate biosensors and bioelectronic devices in situ.
Jia
Wen-Zhi
WZ
Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, PR China.
Wang
Kang
K
Zhu
Zheng-Jiang
ZJ
Song
Hang-Tian
HT
Xia
Xing-Hua
XH
D015374:Biosensing Techniques; D004563:Electrochemistry; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005947:Glucose / Q000737:chemistry; D005949:Glucose Oxidase / Q000737:chemistry*; D006859:Hydrogen / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D044367:Phase Transition; D010984:Platinum / Q000737:chemistry*; D016062:Porosity; D012822:Silicon Dioxide / Q000737:chemistry*
D004800:Enzymes, Immobilized; D010984:Platinum; D012822:Silicon Dioxide; D006859:Hydrogen; D005949:Glucose Oxidase; D005947:Glucose