Çanakkale Onsekiz Mart University, Faculty of Engineering, Bioengineering Department, Çanakkale, Turkey.
Çanakkale Onsekiz Mart University, Faculty of Engineering, Bioengineering Department, Çanakkale, Turkey.
Int J Biol Macromol. 2019 Jun 1;130:245-252. doi: 10.1016/j.ijbiomac.2019.02.112. Epub 2019 Feb 20.
In this study it is aimed to construct an immunosensor system for detection of Sex-determining region Y-box 2 (SOX2) antigen by using Indium tin oxide- polyethylene terephthalate (ITO-PET) as working electrode. Firstly, ITO-PET electrode surfaces were hydroxylated by using NHOH/HO/HO and were incubated with 11-cyanoundecyltrimethoxysilane (11-CUTMS), respectively. After silanization process, anti- SOX2 was immobilized on modified ITO-PET electrodes. All immobilization processes were examined with Electrochemical impedance spectroscopy (EIS) and Cyclic voltammetry (CV). The basic parameters such as 11-CUTMS and anti-SOX2 concentrations, anti-SOX2 and SOX2 incubation period were optimized. The immunosensor prepared under optimal conditions gave a response for a wide concentration range of SOX2 protein (0.02 pg mL-2 pg mL) and the limit of detection was determined as low as 0.013 pg mL. And also, the immunosensor had good repeatability, reproducibility, reusability and long shelf life. Scanning Electron Microscope (SEM) method was utilized to observe the morphologies of the electrode surfaces belonging to all steps. Lastly, seven different real human serum were analyzed with the constructed immunosensor and Enzyme-Linked ImmunoSorbent Assay (ELISA). The results found with these methods were analogised with each other.
本研究旨在构建一种基于氧化铟锡-聚对苯二甲酸乙二醇酯（ITO-PET）作为工作电极的 Sex-determining region Y-box 2（SOX2）抗原免疫传感器系统。首先，通过使用 NHOH/HO/HO 将 ITO-PET 电极表面进行羟化，并分别用 11-氰基十一烷三乙氧基硅烷（11-CUTMS）进行硅烷化处理。在硅烷化处理后，将抗 SOX2 固定在修饰后的 ITO-PET 电极上。所有的固定化过程都通过电化学阻抗谱（EIS）和循环伏安法（CV）进行了检测。优化了 11-CUTMS 和抗 SOX2 浓度、抗 SOX2 和 SOX2 孵育时间等基本参数。在最佳条件下制备的免疫传感器对 SOX2 蛋白的宽浓度范围（0.02 pg/mL-2 pg/mL）有响应，检测限低至 0.013 pg/mL。此外，该免疫传感器具有良好的重复性、重现性、可重复性和长保质期。扫描电子显微镜（SEM）方法用于观察属于所有步骤的电极表面的形貌。最后，用构建的免疫传感器和酶联免疫吸附试验（ELISA）分析了七种不同的人血清。这些方法的结果相互类比。
