一种基于氧化还原探针修饰的电子转移线界面和有效固定化抗体的用于灵敏检测癌胚抗原的无试剂电化学免疫传感器。

A reagentless electrochemical immunosensor for sensitive detection of carcinoembryonic antigen based on the interface with redox probe-modified electron transfer wires and effectively immobilized antibody.

作者信息

Zhang Jing, Yang Luoxing, Pei Jie, Tian Yanzhang, Liu Jiyang

机构信息

Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Third Hospital of Shanxi Medical University, Taiyuan, China.

Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

出版信息

Front Chem. 2022 Aug 8;10:939736. doi: 10.3389/fchem.2022.939736. eCollection 2022.

Abstract

Convenient and sensitive detection of tumors marked in serum samples is of great significance for the early diagnosis of cancers. Facile fabrication of reagentless electrochemical immunosensor with efficient sensing interface and high sensitivity is still a challenge. Herein, an electrochemical immunosensor was easily fabricated based on the easy fabrication of immunoassay interface with electron transfer wires, confined redox probes, and conveniently immobilized antibodies, which can achieve sensitive and reagentless determination of the tumor marker, carcinoembryonic antigen (CEA). Carboxyl multi-walled carbon nanotubes (MWCNTs) were firstly modified with an electrochemical redox probe, methylene blue (MB), which has redox potentials distinguished from those of redox molecules commonly existing in biological samples (for example, ascorbic acid and uric acid). After the as-prepared MB-modified MWCNT (MWCNT-MB) was coated on the supporting glassy carbon electrode (GCE), the MWCNT-MB/GCE exhibited improved active area and electron transfer property. Polydopamine (PDA) was then synthesized through simple self-polymerization of dopamine, which acts as the bio-linker to covalently immobilize the anti-CEA antibody (Ab). The developed immunosensor could be applied for electrochemical detection of CEA based on the decrease in the redox signal of MB after specific binding of CEA and immobilized Ab. The fabricated immunosensor can achieve sensitive determination of CEA ranging from 10 pg/ml to 100 ng/ml with a limit of detection (LOD) of 0.6 pg/ml. Determination of CEA in human serum samples was also realized with high accuracy.

摘要

方便灵敏地检测血清样本中标记的肿瘤对于癌症的早期诊断具有重要意义。制备具有高效传感界面和高灵敏度的无试剂电化学免疫传感器仍然是一项挑战。在此,基于带有电子转移线、受限氧化还原探针和方便固定抗体的免疫分析界面的简便制备,轻松构建了一种电化学免疫传感器,该传感器能够实现对肿瘤标志物癌胚抗原(CEA)的灵敏且无试剂测定。首先用氧化还原探针亚甲基蓝(MB)修饰羧基多壁碳纳米管(MWCNT),其氧化还原电位与生物样本中常见的氧化还原分子(例如抗坏血酸和尿酸)不同。将制备好的MB修饰的MWCNT(MWCNT-MB)涂覆在支撑玻碳电极(GCE)上后,MWCNT-MB/GCE表现出改善的活性面积和电子转移性能。然后通过多巴胺的简单自聚合合成聚多巴胺(PDA),其作为生物连接剂共价固定抗CEA抗体(Ab)。所开发的免疫传感器可基于CEA与固定化Ab特异性结合后MB氧化还原信号的降低用于CEA的电化学检测。所制备的免疫传感器能够实现对10 pg/ml至100 ng/ml范围内CEA的灵敏测定,检测限(LOD)为0.6 pg/ml。还实现了对人血清样本中CEA的高精度测定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07b5/9393226/8767814e233f/fchem-10-939736-g001.jpg

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