Faculty of Pharmacy, Department of Analytical Chemistry, Ege University, 35100, Izmir, Bornova, Turkey.
Graduate School of Natural and Applied Sciences, Department of Biomedical Technologies, Ege University, 35100, Izmir, Bornova, Turkey.
Anal Bioanal Chem. 2020 Nov;412(28):7851-7860. doi: 10.1007/s00216-020-02916-x. Epub 2020 Sep 16.
In the present study, a sensitive electrochemical aptamer-based biosensing strategy for human non-small cell lung cancer (NSCLC) detection was proposed using nanofiber-modified disposable pencil graphite electrodes (PGEs). The composite nanofiber was comprised of polyacrylonitrile (PAN) and polypyrrole (PPy) polymers, and fabrication of the nanofibers was accomplished using electrospinning process onto PGEs. Development of the nanofibers was confirmed using scanning electron microscopy (SEM). The high-affinity 5'-aminohexyl-linked aptamer was immobilized onto a PAN/PPy composite nanofiber-modified sensor surface via covalent bonding strategy. After incubation with NSCLC living cells (A549 cell line) at 37.5 °C, the recognition between aptamer and target cells was monitored by electrochemical impedance spectroscopy (EIS). The selectivity of the aptasensor was evaluated using nonspecific human cervical cancer cells (HeLa) and a nonspecific aptamer sequence. The proposed electrochemical aptasensor showed high sensitivity toward A549 cells with a detection limit of 1.2 × 10 cells/mL. The results indicate that our label-free electrochemical aptasensor has great potential in the design of aptasensors for the diagnostics of other types of cancer cells with broad detection capability in clinical analysis. Graphical abstract.
在本研究中,提出了一种使用纳米纤维修饰的一次性铅笔石墨电极(PGE)的基于适体的灵敏电化学生物传感策略,用于检测人非小细胞肺癌(NSCLC)。该复合纳米纤维由聚丙烯腈(PAN)和聚吡咯(PPy)聚合物组成,并通过静电纺丝工艺在 PGE 上制造纳米纤维。使用扫描电子显微镜(SEM)确认了纳米纤维的形成。高亲和力的 5'-氨基己基连接适体通过共价键合策略固定在 PAN/PPy 复合纳米纤维修饰传感器表面上。在 37.5°C 下与 NSCLC 活细胞(A549 细胞系)孵育后,通过电化学阻抗谱(EIS)监测适体与靶细胞之间的识别。通过使用非特异性人宫颈癌细胞(HeLa)和非特异性适体序列评估适体传感器的选择性。所提出的电化学适体传感器对 A549 细胞具有高灵敏度,检测限为 1.2×10 个细胞/mL。结果表明,我们的无标记电化学适体传感器在设计用于其他类型癌细胞诊断的适体传感器方面具有很大的潜力,具有广泛的临床分析检测能力。