Department of Electrical Engineering, Yale University, New Haven, Connecticut 06520, USA.
Nat Nanotechnol. 2012 May 27;7(6):401-7. doi: 10.1038/nnano.2012.82.
Monitoring the binding affinities and kinetics of protein interactions is important in clinical diagnostics and drug development because such information is used to identify new therapeutic candidates. Surface plasmon resonance is at present the standard method used for such analysis, but this is limited by low sensitivity and low-throughput analysis. Here, we show that silicon nanowire field-effect transistors can be used as biosensors to measure protein-ligand binding affinities and kinetics with sensitivities down to femtomolar concentrations. Based on this sensing mechanism, we develop an analytical model to calibrate the sensor response and quantify the molecular binding affinities of two representative protein-ligand binding pairs. The rate constant of the association and dissociation of the protein-ligand pair is determined by monitoring the reaction kinetics, demonstrating that silicon nanowire field-effect transistors can be readily used as high-throughput biosensors to quantify protein interactions.
监测蛋白质相互作用的结合亲和力和动力学在临床诊断和药物开发中很重要，因为这些信息可用于鉴定新的治疗候选物。目前，表面等离子体共振是用于此类分析的标准方法，但该方法受到灵敏度低和高通量分析的限制。在这里，我们表明硅纳米线场效应晶体管可用作生物传感器，以测量具有低至飞摩尔浓度的蛋白配体结合亲和力和动力学。基于这种传感机制，我们开发了一种分析模型来校准传感器响应并量化两个代表性蛋白-配体结合对的分子结合亲和力。通过监测反应动力学来确定蛋白-配体结合对的缔合和解离速率常数，这表明硅纳米线场效应晶体管可作为高通量生物传感器，用于定量蛋白质相互作用。
