Development of a Bioactive Paper Sensor for Detection of Neurotoxins Using Piezoelectric Inkjet Printing of Sol-Gel-Derived Bioinks
Development of a Bioactive Paper Sensor for Detection of Neurotoxins Using Piezoelectric Inkjet Printing of Sol-Gel-Derived Bioinks
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DOI：
10.1021/ac900660p
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发表时间：
2009-07-01
影响因子：
7.4
通讯作者：
Brennan, John D.
中科院分区：
文献类型：
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作者：
Hossain, S. M. Zakir;Luckham, Roger E.;Brennan, John D.
在不需要复杂的仪器的情况下，人们对新策略的兴趣越来越多。例如，检测神经毒素和有机磷酸盐等乙酸酯（ACHE）抑制剂对神经科学，药物评估，药物开发和环境监测具有影响。具有多种试剂的表面功能化，包括酶和发色剂，是有效开发“量油”或侧向流动生物传感器的关键成分。在本文中，我们描述了一种新型的基于纸张的固相生物传感器，该生物传感器利用生物相容性，酶掺杂的酶的，基于Sol-Gel的墨水的压电喷墨打印来创建比色的传感器条。为此，首先打印了聚乙烯胺（PVAM，捕获阴离子剂），然后通过将酶夹在纸上的两层生物相容性的溶胶 - 凝胶衍生的硅胶层中，通过将酶夹在纸上。通过使用Ellman的比色测定法测量ACHE在纸上的残留活性，并捕获了5-硫代-2-硝基苯甲酸酯（TNB）产品，从而成功地检测了使用该传感器的ACHE抑制剂，包括副毒素和黄曲霉毒素B1。 。该测定法提供了良好的检测极限（二氧氧气，类似于100 nm；黄曲霉毒素B1，类似于30 nm）和快速响应时间（<5 min）。可以通过眼睛或使用数码相机和图像分析软件来实现检测，从而避免需要昂贵且复杂的仪器。我们证明，生物活性纸条可以用作量油尺或基于侧向流动的生物传感器。基于溶胶 - 凝胶的膜的使用产生了一个传感器，该传感器保留了酶活性，并在4度C下存储至少60天后可重现结果，从而使该系统适合在现场存储和使用。
There is an increasing interest in new strategies to rapidly detect analytes of clinical and environmental interest without the need for sophisticated instrumentation. As an example, the detection of acetyleholinesterase (AChE) inhibitors such as neurotoxins and organophosphates has implications for neuroscience, drug assessment, pharmaceutical development, and environmental monitoring. Functionalization of surfaces with multiple reagents, including enzymes and chromogenic reagents, is a critical component for the effective development of "dipstick" or lateral flow biosensors. Herein, we describe a novel paper-based solid-phase biosensor that utilizes piezoelectric inkjet printing of biocompatible, enzyme-doped, sol-gel-based inks to create colorimetric sensor strips. For this purpose, polyvinylamine (PVAm, which captures anionic agents) was first printed and then AChE was overprinted by sandwiching the enzyme within two layers of biocompatible sol-gel-derived silica on paper. AChE inhibitors, including paraoxon and aflatoxin B1, were detected successfully using this sensor by measuring the residual activity of AChE on paper, using Ellman's colorimetric assay, with capture of the 5-thio-2-nitrobenzoate (TNB-) product on the PVAm layer. The assay provided good detection limits (paraoxon, similar to 100 nM; aflatoxin B1,similar to 30 nM) and rapid response times (< 5 min). Detection could be achieved either by eye or using a digital camera and image analysis software, avoiding the need for expensive and sophisticated instrumentation. We demonstrate that the bioactive paper strip can be used either as a dipstick or a lateral flow-based biosensor. The use of sol-gel-based entrapment produced a sensor that retained enzyme activity and gave reproducible results after storage at 4 degrees C for at least 60 days, making the system suitable for storage and use in the field.