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A droplet-based novel approach for viable and low volume consumption surface plasmon resonance bio-sensing inside a polydimethylsiloxane microchip

机译:基于液滴的新颖方法可在聚二甲基硅氧烷微芯片内部进行可行且低耗的表面等离振子共振生物传感

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摘要

Over the course of last two decades, surface plasmon resonance (SPR) has emerged as a viable candidate for label-free detection and characterization for a large pool of biological interactions, ranging from hybridization of oligonucleotides to high throughput drug-screening. Conventional SPR bio-sensing involves a step-response method where the SPR sensorgram in response to a switched sequential flow of analyte and buffer is plotted in real-time and fitted to an exponential curve to extract the associative and dissociative reaction rates. Such measurement schemes involve continuous flow conditions where a substantial reagent volume is consumed and is subject to dispersive mixing at flow switching zones. In this paper, we demonstrate a new plug-train SPR technique in a microfluidic chip that separates and singulates solvent plugs in analyte and buffer by an immiscible air phase. Bio-samples are first discretized within plug droplets with volumes in order of few hundred nanoliters or less followed by pressure-driven transport onto SPR sensing sites of this hydrophobically modified SPR microdevise. The kinetic constants ka and kd for a model protein-small molecule interaction pair are extracted from a plug-train signal and are shown to be in reasonable agreement with our previous reports.
机译:在过去的二十年中,表面等离振子共振(SPR)已成为无标记检测和表征大量生物相互作用(从寡核苷酸杂交到高通量药物筛选)的可行候选物。常规的SPR生物传感涉及一种阶跃响应方法,其中响应于分析物和缓冲液的顺序切换流量实时绘制SPR传感图,并将其拟合到指数曲线以提取缔合和解离反应速率。这样的测量方案涉及连续的流动条件,其中消耗了大量的试剂,并且在流动切换区进行分散混合。在本文中,我们演示了一种在微流控芯片中的新型插拔式SPR技术,该技术可通过不混溶的空气相分离和分离分析物和缓冲液中的溶剂塞。首先将生物样品在体积为几百纳升或更小的塞子液滴中离散化,然后将其压力驱动运输到该疏水改性的SPR微型装置的SPR传感位点。从栓塞序列信号中提取了模型蛋白质-小分子相互作用对的动力学常数ka和kd,结果表明它们与我们先前的报道合理吻合。

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