Jiangsu Co-Innovation Center for Efficient Processing and Utilization of Forest Resources and International Innovation Center for Forest Chemicals and Materials, Nanjing Forestry University, Nanjing 210037, China.
Pharmaceutical Analysis Research Center, Tabriz University of Medical Sciences, Tabriz 5166/15731, Iran.
Biosensors (Basel). 2022 Sep 26;12(10):791. doi: 10.3390/bios12100791.
In this work, 2-AG was successfully detected in human plasma samples using a new sandwich-type electrochemical immune device based on poly--cyclodextrin P(-CD) functionalized with AuNPs-DDT and toluidine blue. The P(-CD) ensured the bioactivity and stability of the immobilized 2-AG antibody by providing a broad surface for the efficient immobilization of the biotinylated antibody. To complete the top section of the immunosensor (reporter), an HRP-conjugated antibody of 2-AG (secondary antibody (Ab2)) was attached to the surface of a glassy carbon electrode (GCE) modified by P(-CD), as well as a primarily biotinylated antibody (Ab1). The biosensor fabrication process was monitored using field-emission scanning electron microscope (FE-SEM) and EDS methods. Using the differential pulse voltammetry technique, the immunosensor was utilized for detection of 2-AG in real samples. The suggested interface increased the surface area, which allowed for the immobilization of a large quantity of anti-2-AG antibody while also improving biocompatibility, stability, and electrical conductivity. Finally, the suggested immunosensor's limit of quantitation was determined to be 0.0078 ng/L, with a linear range of 0.0078 to 1.0 ng/L. The results showed that the suggested bioassay can be utilized for diagnosis of 2-AG in clinical samples as a unique and ultrasensitive electrochemical biodevice.
在这项工作中，成功地使用了一种基于聚环糊精（P-CD）的新型三明治型电化学免疫装置检测了人血浆样品中的 2-AG，该装置用 AuNPs-DDT 和甲苯胺蓝功能化。P-CD 通过提供用于生物素化抗体有效固定的宽表面，确保了固定化 2-AG 抗体的生物活性和稳定性。为了完成免疫传感器（报告器）的顶部部分，将 2-AG 的 HRP 缀合抗体（第二抗体（Ab2））连接到由 P-CD 修饰的玻碳电极（GCE）的表面，以及主要的生物素化抗体（Ab1）。使用场发射扫描电子显微镜（FE-SEM）和 EDS 方法监测生物传感器的制造过程。使用差分脉冲伏安法技术，该免疫传感器用于检测实际样品中的 2-AG。所提出的界面增加了表面积，允许固定大量的抗 2-AG 抗体，同时提高了生物相容性、稳定性和导电性。最后，确定所提出的免疫传感器的定量下限为 0.0078 ng/L，线性范围为 0.0078 至 1.0 ng/L。结果表明，该生物测定法可作为独特且超灵敏的电化学生物器件用于临床样品中 2-AG 的诊断。
