Vural Berfin, ÇaliŞkan Meltem, SezgİntÜrk Mustafa Kemal
Department of Bioengineering, Faculty of Engineering, Çanakkale Onsekiz Mart University, Çanakkale Turkey.
Turk J Chem. 2020 Apr 1;44(2):461-471. doi: 10.3906/kim-1909-53. eCollection 2020.
The aim of this research was to design an electrochemical immunosensor for determination of tumour necrosis factor receptor-associated protein-1(TRAP1) antigen, a heat shock protein linked to tumour necrosis factor. The indium-tin oxide covered polyethylene terephthalate (ITO-PET) electrode surface was cleaned and was prepared for the introduction of hydroxyl groups on its surface by using NH4 OH/H2 O2 /H2 O. As a silanization agent for covalent attachment of anti-TRAP1 on the surface of the ITO working electrode, 3-glycidoxypropyltrimethoxysilane (3-GOPS) was used. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to characterize the immobilization steps. A variety of parameters, 3-GOPS and anti-TRAP1 concentrations, and anti-TRAP1 and TRAP1 incubation durations were optimized. After determining the optimum conditions, characterization studies such as repeatability, reproducibility, regeneration, square wave voltammetry, and single frequency impedance were performed. The electrochemical immunosensor has presented an extremely wide determination range for TRAP1 from 0.1 pg/mL to 100 pg/mL.
本研究的目的是设计一种用于测定肿瘤坏死因子受体相关蛋白-1(TRAP1)抗原的电化学免疫传感器,TRAP1是一种与肿瘤坏死因子相关的热休克蛋白。将覆盖有氧化铟锡的聚对苯二甲酸乙二醇酯(ITO-PET)电极表面清洗干净,并通过使用NH4OH/H2O2/H2O在其表面引入羟基。使用3-缩水甘油氧基丙基三甲氧基硅烷(3-GOPS)作为硅烷化剂,用于将抗TRAP1共价连接到ITO工作电极表面。采用循环伏安法(CV)和电化学阻抗谱(EIS)对固定步骤进行表征。对多种参数进行了优化,包括3-GOPS和抗TRAP1的浓度以及抗TRAP1与TRAP1的孵育时间。在确定最佳条件后,进行了诸如重复性、再现性、再生、方波伏安法和单频阻抗等表征研究。该电化学免疫传感器对TRAP1的测定范围极宽,为0.1 pg/mL至100 pg/mL。