Fabrication of a high-sensitivity electrochemical immunosensor by the oriented immobilization of engineered nanobody on nanofibrous membrane

IF 5.3 2区化学 Q1 CHEMISTRY, ANALYTICAL

Microchimica Acta Pub Date : 2024-10-29 DOI:10.1007/s00604-024-06763-w

Peng Wen, Hao Su, Wen-jia Yin, Jian-cheng Hu, Yu Wang, Jin-yi Yang, Zhi-li Xiao, Zhen-lin Xu, Yu-dong Shen, Hong Wang, Bruce D. Hammock

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Abstract

A new type of label-free electrochemical immunosensor for the high-sensitivity determination of parathion was developed based on the oriented immobilization of nanobody (VHH9) on a gold nanoparticle-loaded polyvinyl alcohol/citric acid nanofiber membrane-modified electrode. The morphology characterization and assembly process of the modified materials were investigated using scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). Under the optimum conditions, the label-free electrochemical immunosensor for parathion exhibited a linear range of 0.0015–6400 ng/mL and a low detection limit of 0.48 pg/mL, the signal response of which was 10 times higher than that of the randomly immobilized VHH9. The immunosensor possessed high selectivity, good repeatability and reusability (keeping above 90% of its initial activity after repeating 8 times), and stability (remaining 90% after 9 weeks of storage). Finally, the average recoveries of parathion from food samples were 93.76–105.73% with the coefficient of variation being 2.65–6.85%, showing good correlation with UPLC (R² = 0.9950). Therefore, our nanobody immobilization protocol is simple and effective and proves the potential to be utilized as a promising candidate for sensing platform.

Graphical Abstract

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通过在纳米纤维膜上定向固定工程纳米抗体制作高灵敏度电化学免疫传感器

在金纳米粒子负载的聚乙烯醇/柠檬酸纳米纤维膜修饰电极上定向固定纳米抗体（VHH9）的基础上，开发了一种高灵敏度测定对硫磷的新型无标记电化学免疫传感器。利用扫描电子显微镜（SEM）和电化学阻抗谱（EIS）研究了改性材料的形态特征和组装过程。在最佳条件下，该对硫磷无标记电化学免疫传感器的线性范围为 0.0015-6400 ng/mL，检出限低至 0.48 pg/mL，其信号响应比随机固定的 VHH9 高 10 倍。该免疫传感器具有高选择性、良好的重复性和可重复性（重复 8 次后仍能保持 90% 以上的初始活性）以及稳定性（储存 9 周后仍能保持 90%）。最后，食品样品中对硫磷的平均回收率为 93.76-105.73%，变异系数为 2.65-6.85%，与 UPLC 的相关性良好（R2 = 0.9950）。因此，我们的纳米抗体固定化方案既简单又有效，证明了其作为传感平台候选物的潜力。

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来源期刊

Microchimica Acta 化学-分析化学

CiteScore

9.80

自引率

5.30%

发文量

410

审稿时长

2.7 months

期刊介绍： As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.