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Surface plasmon resonance biosensor for enzymatic detection of small analytes

机译：表面等离子体共振生物传感器，用于酶检测小分析物

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摘要

Surface plasmon resonance (SPR) biosensing is based on the detection of small changes in the refractive index on a gold surface modified with molecular recognition materials, thus being mostly limited to detecting large molecules. In this paper, we report on a SPR biosensing platform suitable to detect small molecules by making use of the mediator-type enzyme microperoxidase-11 (MP11) in layer-by-layer films. By depositing a top layer of glucose oxidase or uricase, we were able to detect glucose or uric acid with limits of detection of 3.4 and 0.27 mu mol l(-1), respectively. Measurable SPR signals could be achieved because of the changes in polarizability of MP11, as it is oxidized upon interaction with the analyte. Confirmation of this hypothesis was obtained with finite difference time domain simulations, which also allowed us to discard the possible effects from film roughness changes observed in atomic force microscopy images. The main advantage of this mediator-type enzyme approach is in the simplicity of the experimental method that does not require an external potential, unlike similar approaches for SPR biosensing of small molecules. The detection limits reported here were achieved without optimizing the film architecture, and therefore the performance can in principle be further enhanced, while the proposed SPR platform may be extended to any system where hydrogen peroxide is generated in enzymatic reactions.
机译：表面等离子体共振（SPR）生物传感基于对用分子识别材料改性的金表面上的折射率的小变化的检测，从而主要限于检测大分子。在本文中，我们通过在层逐层膜中使用介质型酶微氧化酶-11（MP11）来报告适于检测小分子的SPR生物传感平台。通过沉积顶层葡萄糖氧化酶或尿素酶，我们能够分别检测葡萄糖或尿酸，分别检测3.4和0.27μmL1（-1）。可以通过MP11的极化性的变化来实现可测量的SPR信号，因为它在与分析物的相互作用时被氧化。通过有限差分时域模拟获得了对该假设的确认，这也使我们丢弃在原子力显微镜图像中观察到的膜粗糙度变化的可能效果。这种介质型酶方法的主要优点在于，与小分子的SPR生物溶解的类似方法不同，不需要外部电位的实验方法。这里报道的检测限度是在不优化薄膜架构的情况下实现的，因此原则上可以进一步增强性能，而所提出的SPR平台可以扩展到在酶促反应中产生过氧化氢的任何系统。

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