Construction of effective disposable biosensors for point of care testing of nitrite.
Construction of effective disposable biosensors for point of care testing of nitrite.
复制标题
DOI：
10.1016/j.talanta.2015.04.057
复制
发表时间：
2015-09-01
期刊：
影响因子：
6.1
通讯作者：
Almeida MG
中科院分区：
文献类型：
--
作者：
Monteiro T;Rodrigues PR;Gonçalves AL;Moura JJ;Jubete E;Añorga L;Piknova B;Schechter AN;Silveira CM;Almeida MG
在本文中，我们的目的是证明，作为一个概念验证，大规模生产的有效点的护理测试的亚硝酸盐定量在环境，食品和临床样品的可行性。在我们之前关于基于氨形成亚硝酸盐还原酶（ccNiR）的第三代电化学生物传感器的开发的工作之后，本文中我们将电极系统的尺寸减小到小型化格式，解决了氧干扰的问题，并在真实的样品中进行简单的定量测定。特别地，碳糊丝网印刷电极（SPE）涂覆有在有机溶剂中均质化并在低温下固化的ccNiR/碳墨复合物。通过循环伏安法评估这些化学和热处理的生物相容性，表明丙酮和40 °C固化温度的组合的催化性能更高。蛋白质在碳墨/溶剂复合物中的成功掺入，同时保持催化活性，证明了ccNiR的稳健性和适用于基于丝网印刷的生物传感器的应用。由于分子氧的直接电化学还原发生在电分析测量在激活ccNiR（ca. -0.4 V vs Ag/AgCl），成功地使用了基于葡萄糖氧化酶和过氧化氢酶活性偶联的氧清除系统。这使得不同样品（牛奶、水、血浆和尿液）中的亚硝酸盐定量能够以简单的方式进行，误差很小（1-6%）。在优化的条件下，该传感器对亚硝酸盐还原的灵敏度为0.55 A M−1 cm−2，线性响应范围为0.7-370 μM。
In this paper we aim to demonstrate, as a proof-of-concept, the feasibility of the mass production of effective point of care tests for nitrite quantification in environmental, food and clinical samples. Following our previous work on the development of third generation electrochemical biosensors based on the ammonia forming nitrite reductase (ccNiR), herein we reduced the size of the electrodes’ system to a miniaturized format, solved the problem of oxygen interference and performed simple quantification assays in real samples. In particular, carbon paste screen printed electrodes (SPE) were coated with a ccNiR/carbon ink composite homogenized in organic solvents and cured at low temperatures. The bio-compatibility of these chemical and thermal treatments was evaluated by cyclic voltammetry showing that the catalytic performance was higher with the combination acetone and a 40 °C curing temperature. The successful incorporation of the protein in the carbon ink/solvent composite, while remaining catalytically competent, attests for ccNiR’s robustness and suitability for application in screen printed based biosensors. Because the direct electrochemical reduction of molecular oxygen occurs when electroanalytical measurements are performed at the negative potentials required to activate ccNiR (ca. −0.4 V vs Ag/AgCl), an oxygen scavenging system based on the coupling of glucose oxidase and catalase activities was successfully used. This enabled the quantification of nitrite in different samples (milk, water, plasma and urine) in a straightforward way and with small error (1–6%). The sensitivity of the biosensor towards nitrite reduction under optimized conditions was 0.55 A M−1 cm−2 with a linear response range 0.7–370 μM.
登录
查看更多内容
DOI：
10.1016/j.niox.2009.07.002
发表时间：
2009-09
期刊：
Nitric oxide : biology and chemistry
影响因子：
--
作者：
Hall CN;Garthwaite J
通讯作者：
Garthwaite J
影响因子：
5
作者：
Loaiza, Oscar A.;Lamas-Ardisana, Pedro J.;Grande, Hans J.
通讯作者：
Grande, Hans J.
影响因子：
--
作者：
Dutt, J;Davis, J
通讯作者：
Davis, J
影响因子：
6.2
作者：
Serra, A. S.;Jorge, S. R.;Almeida, M. G.
通讯作者：
Almeida, M. G.
影响因子：
12.6
作者：
Silveira, Celia M.;Gomes, Sofia P.;Almeida, M. Gabriela
通讯作者：
Almeida, M. Gabriela