Karimi A, Othman A, Andreescu S
Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, New York, USA.
Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, New York, USA.
Methods Enzymol. 2016;571:177-95. doi: 10.1016/bs.mie.2016.03.006. Epub 2016 Mar 22.
Portable, nanoparticle (NP)-enhanced enzyme sensors have emerged as powerful devices for qualitative and quantitative analysis of a variety of analytes for biomedicine, environmental applications, and pharmaceutical fields. This chapter describes a method for the fabrication of a portable, paper-based, inexpensive, robust enzyme biosensor for the detection of substrates of oxidase enzymes. The method utilizes redox-active NPs of cerium oxide (CeO2) as a sensing platform which produces color in response to H2O2 generated by the action of oxidase enzymes on their corresponding substrates. This avoids the use of peroxidases which are routinely used in conjunction with glucose oxidase. The CeO2 particles serve dual roles, as high surface area supports to anchor high loadings of the enzyme as well as a color generation reagent, and the particles are recycled multiple times for the reuse of the biosensor. These sensors are small, light, disposable, inexpensive, and they can be mass produced by standard, low-cost printing methods. All reagents needed for the analysis are embedded within the paper matrix, and sensors stored over extended periods of time without performance loss. This novel sensor is a general platform for the in-field detection of analytes that are substrates for oxidase enzymes in clinical, food, and environmental samples.
便携式、纳米颗粒(NP)增强型酶传感器已成为用于生物医学、环境应用和制药领域中各种分析物定性和定量分析的强大设备。本章描述了一种用于制造便携式、纸质、廉价、坚固的酶生物传感器的方法,该传感器用于检测氧化酶的底物。该方法利用氧化铈(CeO2)的氧化还原活性纳米颗粒作为传感平台,其会因氧化酶作用于相应底物产生的过氧化氢而产生颜色变化。这避免了常规与葡萄糖氧化酶一起使用的过氧化物酶的使用。CeO2颗粒具有双重作用,既作为高比表面积载体以锚定高负载量的酶,又作为颜色生成试剂,并且这些颗粒可多次循环使用以实现生物传感器的重复利用。这些传感器体积小、重量轻、一次性使用、成本低,并且可以通过标准的低成本印刷方法进行大规模生产。分析所需的所有试剂都嵌入在纸质基质中,传感器可长时间储存而不会损失性能。这种新型传感器是用于临床、食品和环境样品中作为氧化酶底物的分析物现场检测的通用平台。