"Gleb Wataghin" Physics Institute, University of Campinas , Campinas, São Paulo 13083-859, Brazil.
Kavli Institute of Nanoscience, Delft University of Technology , 2629 HZ Delft, The Netherlands.
Nano Lett. 2017 Oct 11;17(10):5938-5949. doi: 10.1021/acs.nanolett.7b01803. Epub 2017 Sep 19.
Electrically active field-effect transistors (FET) based biosensors are of paramount importance in life science applications, as they offer direct, fast, and highly sensitive label-free detection capabilities of several biomolecules of specific interest. In this work, we report a detailed investigation on surface functionalization and covalent immobilization of biomarkers using biocompatible ethanolamine and poly(ethylene glycol) derivate coatings, as compared to the conventional approaches using silica monoliths, in order to substantially increase both the sensitivity and molecular selectivity of nanowire-based FET biosensor platforms. Quantitative fluorescence, atomic and Kelvin probe force microscopy allowed detailed investigation of the homogeneity and density of immobilized biomarkers on different biofunctionalized surfaces. Significantly enhanced binding specificity, biomarker density, and target biomolecule capture efficiency were thus achieved for DNA as well as for proteins from pathogens. This optimized functionalization methodology was applied to InP nanowires that due to their low surface recombination rates were used as new active transducers for biosensors. The developed devices provide ultrahigh label-free detection sensitivities ∼1 fM for specific DNA sequences, measured via the net change in device electrical resistance. Similar levels of ultrasensitive detection of ∼6 fM were achieved for a Chagas Disease protein marker (IBMP8-1). The developed InP nanowire biosensor provides thus a qualified tool for detection of the chronic infection stage of this disease, leading to improved diagnosis and control of spread. These methodological developments are expected to substantially enhance the chemical robustness, diagnostic reliability, detection sensitivity, and biomarker selectivity for current and future biosensing devices.
基于场效应晶体管 (FET) 的电活性生物传感器在生命科学应用中至关重要,因为它们提供了直接、快速和高度敏感的几种特定生物分子的无标记检测能力。在这项工作中,我们报告了使用生物相容性乙醇胺和聚乙二醇 (PEG) 衍生物涂层进行表面功能化和生物标志物共价固定的详细研究,与使用硅胶整体的传统方法相比,这大大提高了基于纳米线的 FET 生物传感器平台的灵敏度和分子选择性。定量荧光、原子力和 Kelvin 探针力显微镜允许详细研究不同生物功能化表面上固定化生物标志物的均一性和密度。因此,在 DNA 以及病原体蛋白质的情况下,实现了显著增强的结合特异性、生物标志物密度和靶标生物分子捕获效率。这种优化的功能化方法被应用于 InP 纳米线,由于其低表面复合速率,被用作生物传感器的新型有源换能器。所开发的器件提供了超高的无标记检测灵敏度,对于特定的 DNA 序列,通过器件电阻的净变化来测量,灵敏度约为 1 fM。对于一种恰加斯病蛋白标志物 (IBMP8-1),也实现了类似水平的超灵敏检测,灵敏度约为 6 fM。因此,所开发的 InP 纳米线生物传感器为检测这种疾病的慢性感染阶段提供了一种合格的工具,从而改善了诊断和传播控制。这些方法学的发展有望大大提高当前和未来生物传感器设备的化学稳健性、诊断可靠性、检测灵敏度和生物标志物选择性。