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使用基于阵列的酶促光学生物传感器同时测定pH值、尿素、乙酰胆碱和重金属。

Simultaneous determination of pH, urea, acetylcholine and heavy metals using array-based enzymatic optical biosensor.

作者信息

Tsai Hsiao-chung, Doong Ruey-an

机构信息

Department of Atomic Science, National Tsing Hua University, 101, Sec. 2, Kuang Fu Road, Hsinchu 30013, Taiwan.

出版信息

Biosens Bioelectron. 2005 Mar 15;20(9):1796-804. doi: 10.1016/j.bios.2004.07.008.

Abstract

An array-based optical biosensor for the simultaneous analysis of multiple samples in the presence of unrelated multi-analytes was fabricated. Urease and acetylcholinesterase (AChE) were used as model enzymes and were co-entrapped with the sensing probe, FITC-dextran, in the sol-gel matrix to measure pH, urea, acetylcholine (ACh) and heavy metals (enzyme inhibitors). Environmental and biological samples spiked with metal ions were also used to evaluate the application of the array biosensor to real samples. The biosensor exhibited high specificity in identifying multiple analytes. No obvious cross-interference was observed when a 50-spot array biosensor was used for simultaneous analysis of multiple samples in the presence of multiple analytes. The sensing system can determine pH over a dynamic range from 4 to 8.5. The limits of detection (LODs) of 2.5-50 microM with a dynamic range of 2-3 orders of magnitude for urea and ACh measurements were obtained. Moreover, the urease-encapsulated array biosensor was used to detect heavy metals. The analytical ranges of Cd(II), Cu(II), and Hg(II) were between 10 nM and 100 mM. When real samples were spiked with heavy metals, the array biosensor also exhibited potential effectiveness in screening enzyme inhibitors.

摘要

制备了一种基于阵列的光学生物传感器,用于在存在无关多分析物的情况下同时分析多个样品。脲酶和乙酰胆碱酯酶(AChE)用作模型酶,并与传感探针异硫氰酸荧光素 - 葡聚糖共包埋在溶胶 - 凝胶基质中,以测量pH、尿素、乙酰胆碱(ACh)和重金属(酶抑制剂)。添加了金属离子的环境和生物样品也用于评估阵列生物传感器在实际样品中的应用。该生物传感器在识别多种分析物时表现出高特异性。当使用50点阵列生物传感器在存在多种分析物的情况下同时分析多个样品时,未观察到明显的交叉干扰。传感系统可在4至8.5的动态范围内测定pH。尿素和ACh测量的检测限(LOD)为2.5 - 50 microM,动态范围为2 - 3个数量级。此外,包埋脲酶的阵列生物传感器用于检测重金属。Cd(II)、Cu(II)和Hg(II)的分析范围在10 nM至100 mM之间。当实际样品中添加重金属时,阵列生物传感器在筛选酶抑制剂方面也表现出潜在的有效性。

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