Namık Kemal University, Scientific and Technological Research Center, Tekirdağ, Turkey.
Çanakkale 18 Mart University, Faculty of Engineering, Bioengineering Department, Çanakkale, Turkey.
Talanta. 2017 Sep 1;172:162-170. doi: 10.1016/j.talanta.2017.05.048. Epub 2017 May 17.
A novel, sensitive, disposable indium tin oxide (ITO)-based electrochemical immunosensor was developed firstly for simple, rapid determination of Sex-determining region Y-box 2 (SOX2). SOX2 is a cancer biomarker and used for detecting small cell lung cancer, lung adenocarcinoma, squamous cell carcinoma, skin cancer, prostate cancer, and breast cancer. In this study, a disposable ITO thin film based electrode was used as working electrode for biosensing the interaction between SOX2 antigen and anti-SOX2 antibody. In this study, carboxyethylsilanetriol (CTES) was also utilized for electrode modifying so as to obtain self-assembled monolayers. The formed self-assembled monolayers were activated with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)/N-hydroxysuccinimide (NHS) chemistry and they were used as a heterobifunctional crosslinker and activator, respectively. Anti-SOX2 antibody was used as a biorecognition molecule and was covalently immobilized onto the ITO thin film modified with CTES. Immobilization steps were characterized by electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The optimum immobilization conditions such as antibody concentration, antibody and antigen incubation times were examined for the best sensitivity of the immunosensor. Under optimal conditions, this immunosensor had a wide linear detection range (25fg/mL-2pg/mL) with a detection limit as low as 7fg/mL SOX2. Furthermore, the developed SOX2 immunosensor had good storage stability (79.36% of initial activity after 9 weeks), repeatability (3.88% of RSD) and reproducibility (4.25% of RSD). Our developed immunosensor has an acceptable performance for detection of SOX2 antigen, exhibits low detection limit, and has selective and reproducible results in immunoreaction analysis.
一种新型、灵敏、一次性的氧化铟锡（ITO）电化学免疫传感器首次被开发用于简单、快速测定性别决定区 Y 框 2（SOX2）。SOX2 是一种癌症生物标志物，用于检测小细胞肺癌、肺腺癌、鳞状细胞癌、皮肤癌、前列腺癌和乳腺癌。在本研究中，使用一次性 ITO 薄膜作为工作电极用于生物传感 SOX2 抗原与抗 SOX2 抗体之间的相互作用。在本研究中，羧乙基硅烷三醇（CTES）也被用于修饰电极，以获得自组装单层。形成的自组装单层通过 1-乙基-3-（3-二甲基氨基丙基）碳化二亚胺（EDC）/N-羟基琥珀酰亚胺（NHS）化学被激活，它们分别用作异双功能交联剂和激活剂。抗 SOX2 抗体被用作生物识别分子，并通过共价固定在 CTES 修饰的 ITO 薄膜上。通过电化学阻抗谱（EIS）、循环伏安法（CV）、傅里叶变换红外光谱（FTIR）、扫描电子显微镜（SEM）和原子力显微镜（AFM）对固定化步骤进行了表征。检查了最佳固定化条件，如抗体浓度、抗体和抗原孵育时间，以获得免疫传感器的最佳灵敏度。在最佳条件下，该免疫传感器具有宽线性检测范围（25fg/mL-2pg/mL），检测限低至 7fg/mL SOX2。此外，所开发的 SOX2 免疫传感器具有良好的储存稳定性（9 周后初始活性的 79.36%）、重复性（3.88%的 RSD）和再现性（4.25%的 RSD）。我们开发的免疫传感器具有用于检测 SOX2 抗原的可接受性能，表现出低检测限，并在免疫反应分析中具有选择性和可重复性结果。
