Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran; Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Talanta. 2021 Aug 1;230:122276. doi: 10.1016/j.talanta.2021.122276. Epub 2021 Mar 19.
Almost 13% of all death in the world is related to cancer. One of the major reasons for failing cancer treatment is the late diagnosis of the tumors. Thus, diagnosis at the early stages could be vital for the treatment. Serum autoantibodies, as tumor markers, are becoming interesting targets due to their medical and biological relevance. Among them, anti-p53 autoantibody in human sera is found to be involved in a variety of cancers. Regarding this issue, a novel and sensitive electrochemical biosensor for detection of anti-p53 autoantibody has been developed. For this purpose, a nanocomposite including thionine (as an electron transfer mediator)/chitosan/nickel hydroxide nanoparticles/electrochemically reduced graphene oxide (Th-CS-Ni(OH)NPs-ERGO) as a support platform was fabricated on the surface of glassy carbon electrode via a layer-by-layer manner and characterized through common electrochemical and imaging techniques. Then, p53-antigen was immobilized on the nanocomposite and used in an indirect immunoassay with horseradish peroxidase (HRP)-conjugated secondary antibody and HO as the substrate, following the typical Michaelis-Menten kinetics. Under optimized condition, two techniques, including differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS) as a label free technique, applied for the biomarker detection. The linear ranges and LODs were obtained 0.1-500 pg mL and 0.001 pg mL using DPV and 5-150 pg mL and 0.007 pg mL using EIS, respectively. Furthermore, the proposed biosensor displayed satisfying stability, selectivity, and reproducibility. According to the results, the presented protocol is promising to develop other electrochemical biosensors.
全球近 13%的死亡与癌症有关。癌症治疗失败的主要原因之一是肿瘤的晚期诊断。因此,早期诊断对于治疗至关重要。血清自身抗体作为肿瘤标志物,由于其医学和生物学相关性,正成为研究的热点。其中,人血清中的抗 p53 自身抗体被发现与多种癌症有关。关于这个问题,已经开发出了一种用于检测抗 p53 自身抗体的新型灵敏电化学生物传感器。为此,通过层层法在玻碳电极表面制备了包含硫堇(作为电子转移介体)/壳聚糖/氢氧化镍纳米粒子/电化学还原氧化石墨烯(Th-CS-Ni(OH)NPs-ERGO)的纳米复合材料,并通过常见的电化学和成像技术对其进行了表征。然后,将 p53 抗原固定在纳米复合材料上,并与辣根过氧化物酶(HRP)偶联的二抗和 HO 一起用于间接免疫测定,遵循典型的米氏动力学。在优化条件下,两种技术,包括差分脉冲伏安法(DPV)和电化学阻抗谱(EIS)作为无标记技术,用于生物标志物检测。使用 DPV 获得的线性范围和 LOD 分别为 0.1-500 pg mL 和 0.001 pg mL,使用 EIS 获得的线性范围和 LOD 分别为 5-150 pg mL 和 0.007 pg mL。此外,所提出的生物传感器表现出令人满意的稳定性、选择性和重现性。根据结果,所提出的方案有望开发其他电化学生物传感器。