Hashkavayi Ayemeh Bagheri, Raoof Jahan Bakhsh, Ojani Reza
Electroanalytical Chemistry Research Laboratory, Department of Analytical Chemistry, Faculty of Chemistry, University of Mazandaran, Babolsar, 47416-1467, Iran.
Anal Bioanal Chem. 2017 Nov;409(27):6429-6438. doi: 10.1007/s00216-017-0588-z. Epub 2017 Aug 29.
In this work, a highly sensitive, low-cost, and label-free aptasensor based on signal-on mechanisms of response was developed by immobilizing the aptamer on gold nanoparticles (AuNPs)/amine-functionalized silica nanoparticle (FSN)/screen-printed electrode (SPE) surface for highly selective electrochemical detection of tryptophan (Trp). The hemin (Hem), which interacted with the guanine bases of the aptamer, worked as a redox indicator to generate a readable electrochemical signal. The changes in the charge transfer resistance have been monitored using the voltammetry and electrochemical impedance spectroscopic (EIS) techniques. The peak current of Hem linearly increased with increasing concentration of Trp, in differential pulse voltammetry, from 0.06 to 250 nM with a detection limit of 0.026 nM. Also, the results obtained from EIS studies showed that the Trp was detected sensitively with the fabricated aptasensor in the range of 0.06-250 nM. The detection limit is 0.01 nM, much lower than that obtained by most of the reported electrochemical methods. The usage of aptamer as a recognition layer led to a sensor with high affinity for Trp, compared with control amino acids of tyrosine, histidine, arginine, lysine, valine, and methionine. The usability of the aptasensor was successfully evaluated by the determination of Trp in a human blood serum sample. Thus, the sensor could provide a promising plan for the construction of aptasensors. Graphical abstract Schematic outline the principle for tryptophan aptasensing.
在本研究中,通过将适配体固定在金纳米颗粒(AuNPs)/胺功能化二氧化硅纳米颗粒(FSN)/丝网印刷电极(SPE)表面,开发了一种基于信号开启响应机制的高灵敏度、低成本且无标记的适配体传感器,用于高选择性电化学检测色氨酸(Trp)。与适配体鸟嘌呤碱基相互作用的血红素(Hem)作为氧化还原指示剂产生可读取的电化学信号。使用伏安法和电化学阻抗谱(EIS)技术监测电荷转移电阻的变化。在差分脉冲伏安法中,Hem的峰值电流随Trp浓度的增加呈线性增加,范围为0.06至250 nM,检测限为0.026 nM。此外,EIS研究结果表明,所制备的适配体传感器在0.06 - 250 nM范围内能够灵敏地检测Trp。检测限为0.01 nM,远低于大多数已报道的电化学方法。与酪氨酸、组氨酸、精氨酸、赖氨酸、缬氨酸和蛋氨酸等对照氨基酸相比,将适配体用作识别层使得传感器对Trp具有高亲和力。通过测定人血清样品中的Trp成功评估了适配体传感器的实用性。因此,该传感器可为构建适配体传感器提供一个有前景的方案。图形摘要 色氨酸适配体传感原理的示意图。