A novel immobilization multienzyme glucose fluorescence capillary biosensor

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• 作者：Yong-Sheng Li ; Yun-Dong Du ; Ting-Mei Chen ; Xiu-Feng Gao
• 关键词：Fluorescence capillary analysis ; Biosensor ; Immobilization enzyme ; Glucose oxidase ; Horseradish peroxidase ; l-Tyrosine
• 刊名：Biosensors and Bioelectronics
• 出版年：2010
• 出版时间：15 February 2010
• 年：2010
• 卷：25
• 期：6
• 页码：1382-1388
• 全文大小：392 K

文摘

Based on the immobilization enzyme technology and the fluorescence capillary analysis method, the authors have developed a highly sensitive fluorescence reaction system and a novel immobilization multienzyme glucose fluorescence capillary biosensor for determining glucose contents. Reaction principle of the system is that under the catalysis of glucose oxidase (GOD) and horseradish peroxidase (HRP) immobilized on inner surface of a medical capillary, β-d-glucose reacts with dissolved oxygen to form gluconic acid-δ-lactone and hydrogen peroxide, and then the latter reacts with l-tyrosine to produce a tyrosine dimer, which has maximal excitation and emission wavelengths at 320 nm and 410 nm, respectively. Fluorescence of the dimer is proportional to the concentration of the β-d-glucose. Optimization conditions suitable for the reaction system and the biosensor were as follows. Concentration of the l-tyrosine used as fluorescence reagent was 0.15 mol L⁻¹, the active concentrations of the GOD and the HRP for the immobilization were 15 kU L⁻¹ and 8 kU L⁻¹, respectively. Consumptions of the sample and reagents in one determination were 5.0 μL and 15 μL, respectively. Quantitative range of the biosensor for the glucose was in the range 1–10 μmol L⁻¹, its relative standard deviation was less than 4.9 % , and its detection limit was 0.62 μmol L⁻¹. The biosensor's recovery was in the range 96–105 % . Results of some serum determined with the biosensor and with a commercial glucose-kit were well coincident to each other. Accordingly, the biosensor can be applied to the determination of serum glucose contents in the diagnosis of diabetes.