Environment Department, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.
Department of Chemistry, Graduate University of Advanced Technology, Kerman, Iran.
Bioelectrochemistry. 2020 Aug;134:107497. doi: 10.1016/j.bioelechem.2020.107497. Epub 2020 Mar 4.
In the present study, an original electrode fabrication approach was devised to create a label free sensitive electrochemical aptasensor for the detection of Homocysteine (Hcy) (Homocysteine signal was used for detection). To bind certain targets, synthetic oligonucleotides used as aptamers (APs) were specifically selected. Aptamers are substitutes for antibodies for analytical devices because of their sensitivity and high affinity. In this study, Hcy-Binding-Aptamer (HBA) was grafted onto the surface of Au nanoparticles/Glassy Carbon Electrode (Au/GCE) in order to create an aptasensor. The effects of buffer concentration, buffer type, interaction time, and aptamer concentration were investigated and optimized. In addition, Differential Pulse Voltammetry (DPV) was implemented to identify homocysteine. Favorable performance was achieved at a detection limit of 0.01 μM (S/N = 3) and linear range 0.05-20.0 μM. Furthermore, the fabricated aptasensor displayed desirable stability and reproducibility. The developed electrochemical aptasensor was found to have reasonable selectivity for the detection of homocysteine in the presence of cysteine and methionine. Analysis of real samples showed good ability of the proposed homocysteine biosensor to provide sensitive, quick, easy, and cost effective measurement of homocysteine in human blood serum and urine samples.
在本研究中，设计了一种原始的电极制造方法，以创建一种用于检测同型半胱氨酸 (Hcy) 的无标记敏感电化学适体传感器（同型半胱氨酸信号用于检测）。为了结合某些靶标，专门选择了用作适体 (AP) 的合成寡核苷酸。由于其灵敏度和高亲和力，适体可用作分析设备的抗体替代品。在这项研究中，将 Hcy 结合适体 (HBA) 接枝到 Au 纳米粒子/玻璃碳电极 (Au/GCE) 的表面上，以创建适体传感器。研究并优化了缓冲液浓度、缓冲液类型、相互作用时间和适体浓度的影响。此外，还实施了差分脉冲伏安法 (DPV) 以识别同型半胱氨酸。在检测限为 0.01 μM（S/N = 3）和线性范围为 0.05-20.0 μM 时，获得了良好的性能。此外，所制备的适体传感器表现出良好的稳定性和重现性。在存在半胱氨酸和蛋氨酸的情况下，所开发的电化学适体传感器对同型半胱氨酸的检测表现出合理的选择性。对实际样品的分析表明，所提出的同型半胱氨酸生物传感器具有良好的能力，能够灵敏、快速、简便且经济有效地测量人血清和尿液样品中的同型半胱氨酸。
