适体
检出限
微分脉冲伏安法
化学
组合化学
循环伏安法
寡核苷酸
色谱法
胶体金
DNA
电化学
纳米技术
纳米颗粒
电极
生物化学
材料科学
生物
遗传学
物理化学
作者
Hasret Subak,Giulia Selvolini,Marina Macchiagodena,Dilsat Ozkan-Ariksoysal,Marco Pagliai,Piero Procacci,Giovanna Marrazza
标识
DOI:10.1016/j.bioelechem.2020.107691
摘要
This work proposes a voltammetric aptasensor to detect deoxynivalenol (DON) mycotoxin. The development steps of the aptasensor were partnered for the first time to a computational study to gain insights onto the molecular mechanisms involved into the interaction between a thiol-tethered DNA aptamer (80mer-SH) and DON. The exploited docking study allowed to find the binding region of the oligonucleotide sequence and to determine DON preferred orientation. A biotinylated oligonucleotide sequence (20mer-BIO) complementary to the aptamer was chosen to carry out a competitive format. Graphite screen-printed electrodes (GSPEs) were electrochemically modified with polyaniline and gold nanoparticles ([email protected]) by means of cyclic voltammetry (CV) and worked as a scaffold for the immobilization of the DNA aptamer. Solutions containing increasing concentrations of DON and a fixed amount of 20mer-BIO were dropped onto the aptasensor surface: the resulting hybrids were labeled with an alkaline phosphatase (ALP) conjugate to hydrolyze 1-naphthyl phosphate (1-NPP) substrate into 1-naphthol product, detected by differential pulse voltammetry (DPV). According to its competitive format, the aptasensor response was signal-off in the range 5.0–30.0 ng·mL−1 DON. A detection limit of 3.2 ng·mL−1 was achieved within a 1-hour detection time. Preliminary experiments on maize flour samples spiked with DON yielded good recovery values.
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