Protein binding kinetics quantification via coupled plasmonic-photonic resonance nanosensors in generic microplate reader
Protein binding kinetics quantification via coupled plasmonic-photonic resonance nanosensors in generic microplate reader
复制标题
通过通用酶标仪中耦合等离子体光子共振纳米传感器进行蛋白质结合动力学定量
DOI：
10.1016/j.bios.2019.111494
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发表时间：
2019-10-01
影响因子：
12.6
通讯作者：
Liu, Gang Logan
中科院分区：
文献类型：
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作者：
Dang, Tang;Hu, Wenjun;Liu, Gang Logan
对于普通微板阅读器，几乎没有一种分析分析方法，无论是比色分析还是荧光分析，都不能动态测量蛋白质与蛋白质的结合或量化蛋白质相互作用的动力学缔合常数和解离常数。另一方面，蛋白质结合动力学定量可以在特殊昂贵的表面等离子体共振(SPR)传感设备上独一无二地完成。在这里，我们报道了标准96孔板格式的耦合等离子体-光子共振纳米传感器的集成，并首次使用它在通用微板阅读器上演示了无标记的动态SPR类蛋白质结合测量和动力学定量。我们的低成本无标记纳米传感器板能够基于在通用微板阅读器中测量的特定波长下的透射光密度(OD)值变化，非常灵敏地检测固定化蛋白质相互作用。相对终点OD值变化与蛋白质浓度(0.05~50微克/毫升)呈良好的线性关系。血清中蛋白质的定量结果与同时进行的医院实验室检测结果一致。最重要的是，我们的传感器板井中蛋白质相互作用的动力学缔合和解离常数是通过在通用微板读取器中的时移动态OD值测量来确定的。在我们独特的纳米传感器平板的支持下，现在可以使用许多化学和生物医学研究实验室中普遍存在的通用微板读取器来进行类似SPR的蛋白质结合动力学测量。
Almost no analytical assays, either colorimetric or fluorescence assays, for generic microplate readers is capable of dynamic measurements of protein-protein binding or the quantification of kinetic association and dissociation constants of protein interactions. On the other hand, protein binding kinetics quantification can be uniquely done on special expensive surface plasmon resonance (SPR) sensing equipment. Here we report the integration of coupled plasmonic-photonic resonance nanosensors in standard 96-well plate format and by using which, for the very first time, the demonstration of label-free dynamic SPR-like protein binding measurement and kinetics quantification in a generic microplate reader. Our low-cost label-free nanosensor plate enables very sensitive detection of immobilized protein interactions based on the transmission optical density (OD) value changes at specific wavelengths measured in a generic microplate reader. The relative end-point OD value changes show a good linear response with protein concentrations (from 0.05 to 50 mu g/ml). And the protein quantification in serum results are consistent with the concurrent hospital lab tests. Most importantly, the kinetic association and dissociation constants of protein interactions in our sensor plate wells are determined by time-lapse dynamic OD value measurement in the generic microplate reader. Enabled by our unique nanosensor plate, SPR-like measurement of protein binding kinetics is now available using generic microplate reader ubiquitous in many chemistry and biomedical research labs.