Polydopamine Thin Films as Protein Linker Layer for Sensitive Detection of Interleukin-6 by Surface Plasmon Enhanced Fluorescence Spectroscopy
Polydopamine Thin Films as Protein Linker Layer for Sensitive Detection of Interleukin-6 by Surface Plasmon Enhanced Fluorescence Spectroscopy
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DOI：
10.1021/acsami.6b06917
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发表时间：
2016-08-31
影响因子：
9.5
通讯作者：
Tawa, Keiko
中科院分区：
文献类型：
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作者：
Toma, Mana;Tawa, Keiko
利用表面等离子体增强荧光光谱法（SPFS）将聚多巴胺（PDA）薄膜作为捕获抗体的连接层引入到传感器表面的表面修饰中。捕获抗体可以直接连接到传感器表面，而不使用任何偶联剂，通过用PDA薄膜功能化的金传感器表面。PDA涂层通过在传感器表面上施加多巴胺溶液的单步制备过程进行，这需要极短的孵育时间（10分钟）。通过表面等离子体共振（SPR）光谱研究了捕获抗体在PDA涂层传感器表面上的吸附动力学的实时原位测量。其揭示了在引入含有捕获抗体的溶液之后立即发生捕获抗体的固定，并且传感器表面完全被捕获抗体覆盖。通过SPFS使用荧光标记检测抗体的夹心测定形式进行细胞因子标志物白细胞介素-6（IL-6）的灵敏检测。通过PDA化学功能化的传感器芯片表现出灵敏的传感器响应，检测抗体在传感器表面上的非特异性吸附低。所开发的SPFS生物传感器的IL-6的检测限被确定为2 pg/mL（100 fM），这是在诊断标准的范围内。我们的观察阐明了PDA涂层的显着效用的金属传感器表面的化学改性的一个简单的，简短的，和廉价的方式。
Polydopamine (PDA) thin films are introduced to the surface modification of biosensor surfaces utilizing surface plasmon enhanced fluorescence spectroscopy (SPFS) as the linker layer of capture antibody on to the sensor surfaces. The capture antibody can be directly attached to the sensor surface without using any coupling agent by functionalizing the gold sensor surface with PDA thin films. The PDA coating is performed by a single-step preparation process by applying the dopamine solution on the sensor surface, which requires an extremely short incubation time (10 min). The real-time in situ measurement of the adsorption kinetics of the capture antibody onto the PDA-coated sensor surface is studied by surface plasmon resonance (SPR) spectroscopy. It reveals that the immobilization of capture antibody immediately occurs after introduction of a solution containing capture antibody, and the sensor surface is fully covered with the capture antibody. The sensitive detection of the cytokine marker interleukin-6 (IL-6) is performed by SPFS using a sandwich assay format with fluorescently labeled detection antibody. The sensor chips functionalized by PDA chemistry exhibited sensitive sensor responses with low nonspecific adsorption of the detection antibody onto the sensor surface. The detection limit of IL-6 with the developed SPFS biosensor is determined to be 2 pg/mL (100 fM), which is within the range of the diagnostic criteria. Our observation elucidates the remarkable utility of PDA coatings for chemical modification of the metallic sensor surfaces by a simple, brief, and inexpensive manner.