Food and Drug Safety Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Biochemistry, Higher Education Institute of Rab-Rashid, Tabriz, Iran.
J Mol Recognit. 2020 Oct;33(10):e2852. doi: 10.1002/jmr.2852. Epub 2020 Apr 17.
Carcinoembryonic antigen (CEA) is a member of a family of cell surface glycoproteins. Recognition of CEA is needed to monitor the physiological status of the patient for treatment and also it is important to assess the severity of the disease. In this work, we reported a novel sandwich-type electrochemical immunosensor based on gold nanoparticles functionalized cysteamine-glutaraldehyde (AuNPs-CysA-GA) and it successfully designed to detection of the CEA biomarker in a human plasma sample. The AuNPs-CysA-GA provides a large surface area for the effective immobilization of CEA antibody, as well as it ascertains the bioactivity and stability of immobilized CEA antigens. Biotinylated-anti-CEA antibody (Ab1) was immobilized on the surface of glassy carbon electrode (GCE) modified AuNPs-CysA-GA. Also, secondary antibody (HRP-Ab2) was costed immobilized to complete the sandwich part of immunosensor. Field emission scanning electron microscope (FE-SEM and EDS), was employed to monitor the sensor fabrication procedure. The immunosensor was used for the detection of CEA using differential pulse voltammetry (DPVs) technique. The proposed interface led to enhancement of accessible surface area for immobilizing high amount of anti-CEA antibody, increasing electrical conductivity, boosting stability, and biocompatibility. Finally, the low limit of quantitation (LLOQ) of the proposed immunosensor was obtained as 7 ng/mL with the linear range of 0.001-5 μg/L. The proposed immunoassay was successfully applied for the monitoring of the CEA in unprocessed human plasma samples. Obtained results paved that the proposed bioassay can be used as a novel bioassay for the clinical diagnosis of cancer based on CEA monitoring.
癌胚抗原（CEA）是细胞表面糖蛋白家族的一员。识别 CEA 对于监测患者的生理状态以进行治疗是必要的，而且评估疾病的严重程度也很重要。在这项工作中，我们报告了一种基于金纳米粒子功能化半胱氨酸-戊二醛（AuNPs-CysA-GA）的新型夹心型电化学免疫传感器，并成功设计用于检测人血浆样品中的 CEA 生物标志物。AuNPs-CysA-GA 提供了一个大的表面积，用于有效固定 CEA 抗体，并且还确保了固定化 CEA 抗原的生物活性和稳定性。生物素化的抗 CEA 抗体（Ab1）固定在修饰有 AuNPs-CysA-GA 的玻碳电极（GCE）表面上。此外，将第二抗体（HRP-Ab2）固定在表面上以完成免疫传感器的夹心部分。场发射扫描电子显微镜（FE-SEM 和 EDS）用于监测传感器的制造过程。使用差分脉冲伏安法（DPVs）技术检测 CEA 。该传感器用于通过差分脉冲伏安法（DPVs）技术检测 CEA。所提出的界面导致用于固定大量抗 CEA 抗体的可及表面积增加，电导率提高，稳定性和生物相容性提高。最后，获得的建议免疫传感器的最低定量限（LLOQ）为 7ng/mL，线性范围为 0.001-5μg/L。该免疫分析成功地应用于未经处理的人血浆样品中 CEA 的监测。获得的结果表明，所提出的生物测定法可以用作基于 CEA 监测的癌症临床诊断的新型生物测定法。
