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基于电合成金纳米树突/壳聚糖纳米粒子有效平台的新型高灵敏度和选择性肉毒毒素 A 型免疫传感器的制作。

Fabrication of a Novel Highly Sensitive and Selective Immunosensor for Botulinum Neurotoxin Serotype A Based on an Effective Platform of Electrosynthesized Gold Nanodendrites/Chitosan Nanoparticles.

机构信息

Department of Microbiology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran 1435116471, Iran.

Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences, Tehran 1435116471, Iran.

出版信息

Sensors (Basel). 2017 May 9;17(5):1074. doi: 10.3390/s17051074.

Abstract

In this work, a novel nanocomposite consisting of electrosynthesized gold nanodendrites and chitosan nanoparticles (AuNDs/CSNPs) has been prepared to fabricate an impedimetric immunosensor based on a screen printed carbon electrode (SPCE) for the rapid and sensitive immunoassay of botulinum neurotoxin A (BoNT/A). BoNT/A polyclonal antibody was immobilized on the nanocomposite-modified SPCE for the signal amplification. The structure of the prepared nanocomposite was investigated by transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). The charge transfer resistance (R) changes were used to detect BoNT/A as the specific immuno-interactions at the immunosensor surface that efficiently limited the electron transfer of Fe(CN)₆ as a redox probe at pH = 7.4. A linear relationship was observed between the %∆ and the concentration logarithm of BoNT/A within the range of 0.2 to 230 pg·mL with a detection limit (S/N = 3) of 0.15 pg·mL. The practical applicability of the proposed sensor was examined by evaluating the detection of BoNT/A in milk and serum samples with satisfactory recoveries. Therefore, the prepared immunosensor holds great promise for the fast, simple and sensitive detection of BoNT/A in various real samples.

摘要

在这项工作中,制备了一种由电合成的金纳米树突和壳聚糖纳米粒子(AuNDs/CSNPs)组成的新型纳米复合材料,用于基于丝网印刷碳电极(SPCE)的快速灵敏免疫传感器的制备,用于检测肉毒神经毒素 A(BoNT/A)。BoNT/A 多克隆抗体固定在纳米复合材料修饰的 SPCE 上以进行信号放大。通过透射电子显微镜(TEM)、扫描电子显微镜(SEM)、X 射线衍射(XRD)、傅里叶变换红外(FTIR)光谱、循环伏安法(CV)和电化学阻抗谱(EIS)研究了制备的纳米复合材料的结构。通过在 pH = 7.4 时在免疫传感器表面上发生的特定免疫相互作用来检测 BoNT/A,该相互作用有效地限制了作为氧化还原探针的 Fe(CN)₆的电子转移,从而改变了电荷转移电阻(R)变化。在 0.2 至 230 pg·mL 的范围内,观察到 %∆与 BoNT/A 浓度对数之间存在线性关系,检测限(S/N = 3)为 0.15 pg·mL。通过评估牛奶和血清样品中 BoNT/A 的检测,检验了所提出传感器的实际适用性,结果令人满意。因此,所制备的免疫传感器在各种实际样品中快速、简单、灵敏地检测 BoNT/A 具有很大的应用前景。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7528/5470464/ab7f994ba96d/sensors-17-01074-sch001.jpg

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