Coffee ring aptasensor for rapid protein detection.
Coffee ring aptasensor for rapid protein detection.
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DOI：
10.1021/la400224a
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发表时间：
2013-07-02
期刊：
影响因子：
--
通讯作者：
Lillehoj PB
中科院分区：
文献类型：
--
作者：
Wen JT;Ho CM;Lillehoj PB
我们介绍了一种新的生物传感平台，用于快速蛋白质检测，该平台结合了生物分子浓缩的最简单方法之一，咖啡环形成，以及基于灵敏适体的光学检测方案。在该方法中，适体信标用于信号转导，其中在靶分子存在下发射荧光信号。信号放大是通过在液滴内浓缩适体-靶复合物来实现的，导致咖啡环“斑点”的形成。利用具有各种化学涂层的表面来研究表面疏水性、浓缩效率和信号放大之间的相关性。基于我们的研究结果，我们发现，咖啡环直径的增加与较大的液滴体积是独立的表面疏水性。此外，我们表明，高度疏水表面产生增强的颗粒浓度，通过咖啡环的形成，导致信号强度6倍以上的亲水表面。为了验证该生物传感平台用于检测临床样品，我们检测了人血清和4倍稀释的全血中的α-凝血酶。根据我们的研究结果，咖啡环斑点产生的检测信号比液滴中的样品大40倍。此外，该生物传感器在血清中的检测下限为2 ng/mL（54 pM），在血液中的检测下限为4 ng/mL（105 pM）。基于其简单性和高性能，该平台显示出作为检测疾病生物标志物的廉价诊断工具的巨大潜力，特别是用于缺乏常规生物检测实践所需的资源和设施的发展中国家。
We introduce a new biosensing platform for rapid protein detection that combines one of the simplest methods for biomolecular concentration, coffee ring formation, with a sensitive aptamer-based optical detection scheme. In this approach, aptamer beacons are utilized for signal transduction where a fluorescence signal is emitted in the presence of the target molecule. Signal amplification is achieved by concentrating aptamer-target complexes within liquid droplets, resulting in the formation of coffee ring “spots”. Surfaces with various chemical coatings were utilized to investigate the correlation between surface hydrophobicity, concentration efficiency and signal amplification. Based on our results, we found that the increase in coffee ring diameter with larger droplet volumes is independent of surface hydrophobicity. Furthermore, we show that highly hydrophobic surfaces produce enhanced particle concentration, via coffee ring formation, resulting in signal intensities 6-fold greater than those on hydrophilic surfaces. To validate this biosensing platform for the detection of clinical samples, we detected α-thrombin in human serum and 4x diluted whole blood. Based on our results, coffee ring spots produced detection signals 40x larger than samples in liquid droplets. Additionally, this biosensor exhibits a lower limit of detection of 2 ng/mL (54 pM) in serum, and 4 ng/mL (105 pM) in blood. Based on its simplicity and high performance, this platform demonstrates immense potential as an inexpensive diagnostic tool for the detection of disease biomarkers, particularly for use in developing countries that lack the resources and facilities required for conventional biodetection practices.
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