Kumar Suveen, Panwar Shweta, Kumar Saurabh, Augustine Shine, Malhotra Bansi D
Nanobioelectronics Laboratory, Department of Biotechnology, Delhi Technological University, Delhi110042, India.
Department of Chemistry, University of Delhi, Delhi110007, India.
Nanomaterials (Basel). 2019 Aug 22;9(9):1190. doi: 10.3390/nano9091190.
We report results of the studies relating to the development of an efficient biosensor for non-invasive detection of CYFRA-21-1 cancer biomarker. We used a low dielectric constant material (nanostructured yttrium oxide, nYO) for the fabrication of the biosensing platform. The nYO was synthesized via solvothermal process and functionalized using 3-aminopropyl triethoxy silane (APTES). Electrophoretic deposition (EPD) of the functionalized nanomaterial (APTES/nYO) onto an indium tin oxide (ITO)-coated glass electrode was conducted at a DC potential of 50V for 60s. The EDC-NHS chemistry was used for covalent immobilization of -COOH bearing monoclonal anti-CYFRA-21-1 onto -NH groups of APTES/nYO/ITO electrode. To avoid the non-specific interaction on the anti-CYFRA-21-1/APTES/nYO/ITO immunoelectrode, bovine serum albumin (BSA) was used. X-ray diffraction (XRD), transmission electron microscopy (TEM), and field emission scanning electron microscopy (FESEM) were utilized for structural and morphological studies, whereas Fourier-transform infrared spectroscopy (FTIR) was used for the bonding analysis. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques were used for electrochemical characterization and response studies of fabricated electrodes. The fabricated immunosensor (BSA/anti-CYFRA-21-1/APTES/nYO/ITO) exhibited linearity in the range of 0.01-50 ng·mL, sensitivity of 226.0 Ω·mL·ng, and lower detection limit of 0.01·ng·mL. A reasonable correlation was observed between the results obtained using this biosensor and concentration of CYFRA-21-1 measured through ELISA (enzyme-linked immunosorbent assay) technique in salivary samples of oral cancer patients.
我们报告了有关开发用于非侵入性检测CYFRA-21-1癌症生物标志物的高效生物传感器的研究结果。我们使用低介电常数材料（纳米结构氧化钇，nYO）制造生物传感平台。通过溶剂热法合成nYO，并使用3-氨丙基三乙氧基硅烷（APTES）进行功能化。在50V直流电势下进行60秒，将功能化纳米材料（APTES/nYO）电泳沉积到氧化铟锡（ITO）涂层的玻璃电极上。使用EDC-NHS化学方法将带有-COOH的抗CYFRA-21-1单克隆抗体共价固定到APTES/nYO/ITO电极的-NH基团上。为避免抗CYFRA-21-1/APTES/nYO/ITO免疫电极上的非特异性相互作用，使用了牛血清白蛋白（BSA）。利用X射线衍射（XRD）、透射电子显微镜（TEM）和场发射扫描电子显微镜（FESEM）进行结构和形态研究，而傅里叶变换红外光谱（FTIR）用于键合分析。使用循环伏安法（CV）和电化学阻抗谱（EIS）技术对制备的电极进行电化学表征和响应研究。制备的免疫传感器（BSA/抗CYFRA-21-1/APTES/nYO/ITO）在0.01-50 ng·mL范围内表现出线性，灵敏度为226.0 Ω·mL·ng，检测下限为0.01·ng·mL。在使用该生物传感器获得的结果与通过酶联免疫吸附测定（ELISA）技术在口腔癌患者唾液样本中测得的CYFRA-21-1浓度之间观察到合理的相关性。
