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Optimization of a Paper-Based ELISA for a Human Performance Biomarker

机译:基于纸质ELISA的人类性能生物标记物的优化

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Monitoring aspects of human performance during various activities has recently become a highly investigated research area. Many new commercial products are available now to monitor human physical activity or responses while performing activities ranging from playing sports, to driving, and even sleeping. However, monitoring cognitive performance biomarkers, such as neuropeptides, is still an emerging field due to the complicated sample collection and processing, as well as the need for a clinical lab to perform analysis. Enzyme-linked immunosorbent assays (ELISAs) provide specific detection of biomolecules with high sensitivity (picomolar concentrations). Even with the advantage of high sensitivity, most ELISAs need to be performed in a laboratory setting and require around 6 h to complete. Transitioning this assay to a platform where it reduces cost, shortens assay time, and is able to be performed outside a lab is invaluable. Recently developed paper diagnostics provide an inexpensive platform on which to perform ELISAs; however, the major limiting factor for moving out of the laboratory environment is the measurement and analysis instrumentation. Using something as simple as a digital camera or camera-enabled Windows- or Android-based tablets, we are able to image paper-based ELISAs (P-ELISAs), perform image analysis, and produce response curves with high correlation to target biomolecule concentration in the 10 pM range. Neuropeptide Y detection was performed. Additionally, silver enhancement of Au NPs conjugated with IgG antibodies showed a concentration-dependent response to IgG, thus eliminating the need for an enzyme--substrate system. Automated image analysis and quantification of antigen concentrations are able to be performed on Windows- and Android-based mobile platforms.
机译:在各种活动中监视人类绩效的各个方面最近已成为受到高度研究的研究领域。现在,有许多新的商业产品可用来监视人类的身体活动或反应,同时执行从运动到开车甚至睡觉的活动。然而,由于复杂的样品收集和处理以及需要临床实验室进行分析,监测诸如神经肽的认知性能生物标记物仍是新兴领域。酶联免疫吸附测定(ELISA)提供高灵敏度(皮摩尔浓度)的生物分子特异性检测。即使具有高灵敏度的优势,大多数ELISA也需要在实验室环境中进行,大约需要6小时才能完成。将该测定法转移到可降低成本,缩短测定时间并能够在实验室外进行的平台是无价的。最近开发的纸质诊断提供了一种廉价的平台,可以在其上进行ELISA。但是,移出实验室环境的主要限制因素是测量和分析仪器。使用像数码相机或基于相机的基于Windows或Android的平板电脑之类的简单工具,我们就可以对基于纸张的ELISA(P-ELISA)进行成像,执行图像分析并生成与目标生物分子浓度高度相关的响应曲线在10 pM范围内。进行神经肽Y检测。此外,与IgG抗体偶联的Au NP的银增强显示出对IgG的浓度依赖性反应,因此消除了对酶-底物系统的需求。可以在基于Windows和Android的移动平台上执行自动图像分析和抗原浓度定量。

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