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基于三明治结构的超灵敏电化学适体传感器用于选择性无标记检测结直肠癌(CT26)细胞。

Ultrasensitive electrochemical aptasensor based on sandwich architecture for selective label-free detection of colorectal cancer (CT26) cells.

机构信息

Electroanalytical Chemistry Research Laboratory, Department of Analytical Chemistry, Faculty of Chemistry, University of Mazandaran, Babolsar, Iran.

Electroanalytical Chemistry Research Laboratory, Department of Analytical Chemistry, Faculty of Chemistry, University of Mazandaran, Babolsar, Iran.

出版信息

Biosens Bioelectron. 2017 Jun 15;92:630-637. doi: 10.1016/j.bios.2016.10.042. Epub 2016 Oct 19.

Abstract

Colorectal cancer is one of the most common cancers in the world and has no effective treatment. Therefore, development of new methods for early diagnosis is instantly required. Biological recognition probes such as synthetic receptor and aptamer is one of the candidate recognition layers to detect important biomolecules. In this work, an electrochemical aptasensor was developed by fabricating an aptamer-cell-aptamer sandwich architecture on an SBA-15-3-aminopropyltriethoxysilane (SBA-15-pr-NH) and Au nanoparticles (AuNPs) modified graphite screen printed electrode (GSPE) surface for the selective, label-free detection of CT26 cancer cells. Based on the incubation of the thiolated aptamer with CT26 cells, the electron-transfer resistance of Fe (CN) redox couple increased considerably on the aptasensor surface. The results obtained from cyclic voltammetry and electrochemical impedance spectroscopy studies showed that the fabricated aptasensor can specifically identify CT26 cells in the concentration ranges of 10-1.0×10cells/mL and 1.0×10-6.0×10 cells/mL, respectively, with a detection limit of 2cells/mL. Applying the thiol terminated aptamer (5TR1) as a recognition layer led to a sensor with high affinity for CT26 cancer cells, compared to control cancer cells of AGS cells, VERO Cells, PC3 cells and SKOV-3 cells. Therefore a simple, rapid, label free, inexpensive, excellent, sensitive and selective electrochemical aptasensor based on sandwich architecture was developed for detection of CT26 Cells.

摘要

结直肠癌是世界上最常见的癌症之一,目前尚无有效的治疗方法。因此,急需开发新的方法来进行早期诊断。生物识别探针,如合成受体和适体,是用于检测重要生物分子的候选识别层之一。在这项工作中,通过在 SBA-15-3-氨丙基三乙氧基硅烷(SBA-15-pr-NH)和金纳米粒子(AuNPs)修饰的石墨丝网印刷电极(GSPE)表面上构建适体-细胞-适体三明治结构,开发了一种电化学适体传感器,用于选择性、无标记检测 CT26 癌细胞。基于巯基化适体与 CT26 细胞的孵育,Fe(CN)氧化还原对的电子转移电阻在适体传感器表面上显著增加。循环伏安法和电化学阻抗谱研究的结果表明,所制备的适体传感器能够分别在 10-1.0×10^cells/mL 和 1.0×10^-6.0×10^cells/mL 的浓度范围内特异性识别 CT26 细胞,检测限为 2cells/mL。与 AGS 细胞、VERO 细胞、PC3 细胞和 SKOV-3 细胞等对照癌细胞相比,应用硫醇末端适体(5TR1)作为识别层,使传感器对 CT26 癌细胞具有更高的亲和力。因此,开发了一种基于三明治结构的简单、快速、无标记、廉价、优异、灵敏和选择性的电化学适体传感器,用于 CT26 细胞的检测。

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