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Highly sensitive multipoint real-time kinetic detection of Surface Plasmon bioanalytes with custom CMOS cameras

机译:使用定制的CMOS相机对表面等离子生物分析物进行高灵敏度的多点实时动力学检测

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摘要

Phase sensitive Surface Plasmon Resonance (SPR) techniques are a popular means of characterizing biomolecular interactions. However, limitations due to the narrow dynamic range and difficulty in adapting the method for multi-point sensing have restricted its range of applications. This paper presents a compact phase sensitive SPR technology using a custom CMOS camera. The system is exceptionally versatile enabling one to trade dynamic range for sensitivity without altering the optical system. We present results showing sensitivity over the array of better than 10−6 Refractive Index Units (RIU) over a refractive index range of 2×10−2 RIU, with peak sensitivity of 3×10−7 RIU at the center of this range. We also explain how simply altering the settings of polarization components can give sensitivity on the order of 10−8 RIU albeit at the cost of lower dynamic range. The consistent response of the custom CMOS camera in the system also allowed us to demonstrate precise quantitative detection of two Fibrinogen antibody–protein binding sites. Moreover, we use the system to determine reaction kinetics and argue how the multipoint detection gives useful insight into the molecular binding mechanisms.
机译:相敏表面等离子体共振(SPR)技术是表征生物分子相互作用的一种流行方法。但是,由于动态范围狭窄以及难以适应多点传感方法的限制,限制了其应用范围。本文介绍了一种使用定制CMOS相机的紧凑型相敏SPR技术。该系统具有出色的通用性,可以在不改变光学系​​统的情况下以动态范围进行灵敏度交换。我们提供的结果显示,在2×10 -2 RIU的折射率范围内,优于10 −6 折射率单位(RIU)的阵列的灵敏度为峰值在此范围中心的3×10 −7 RIU我们还解释了如何简单地改变偏振分量的设置,尽管以降低动态范围为代价,却可以使灵敏度达到10 -8 RIU的数量级。系统中定制CMOS相机的一致响应也使我们能够证明对两个纤维蛋白原抗体-蛋白质结合位点的精确定量检测。此外,我们使用该系统确定反应动力学,并论证了多点检测如何对分子结合机理提供有用的见解。

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