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基于电化学介导原子转移自由基聚合的细胞角蛋白 21-1 电化学发光免疫传感器的构建

Electrochemiluminescence immunosensor for cytokeratin fragment antigen 21-1 detection using electrochemically mediated atom transfer radical polymerization.

机构信息

Pharmacy College, Henan University of Chinese Medicine, Zhengzhou, 450046, People's Republic of China.

The Engineering and Technology Center for Chinese Medicine Development of Henan Province, Zhengzhou, 450046, People's Republic of China.

出版信息

Mikrochim Acta. 2021 Mar 8;188(4):115. doi: 10.1007/s00604-020-04677-x.

Abstract

The cytokeratin fragment antigen 21-1 (CYFRA 21-1) protein is a critical tumor biomarker tightly related to non-small cell lung cancer (NSCLC). Herein, we prepared an effective electrochemiluminescence (ECL) immunosensor for CYFRA 21-1 detection using electrochemically mediated atom transfer radical polymerization (eATRP). The CYFRA 21-1 antigen was fixed on the electrode surface by constructing a sandwich type antibody-antigen-antibody immune system. The sensitivity of ECL was improved by using the eATRP reaction. In this method, eATRP was applied to CYFRA 21-1 detection antibody with N-acryloyloxysuccinimide as functional monomer. This is the first time that ECL and eATRP signal amplification technology had been combined. Under the optimized testing conditions, the immunosensor showed a good linear relation in the range from 1 fg mL to 1 μg mL at a limit of detection of 0.8 fg mL (equivalent to ~ 134 molecules in a 10 μL sample). The ECL immunosensing system based on eATRP signal amplification technology provided a new way for rapid diagnosis of lung cancer by detecting CYFRA 21-1. The paper prepared an electrochemiluminescence biosensor for ultrasensitive detection of CYFRA 21-1 via eATRP signal amplification strategy, which had the advantages of high sensitivity, reproducibility, and held potential prospect for analysis of low-abundance.

摘要

细胞角蛋白 21-1(CYFRA 21-1)蛋白是一种与非小细胞肺癌(NSCLC)密切相关的关键肿瘤标志物。在此,我们使用电化学介导的原子转移自由基聚合(eATRP)制备了一种用于检测 CYFRA 21-1 的有效电化学发光(ECL)免疫传感器。通过构建三明治型抗体-抗原-抗体免疫体系,将 CYFRA 21-1 抗原固定在电极表面。通过 eATRP 反应提高了 ECL 的灵敏度。在该方法中,将 eATRP 应用于 CYFRA 21-1 检测抗体,其中 N-丙烯酰氧基琥珀酰亚胺作为功能单体。这是首次将 ECL 和 eATRP 信号放大技术结合使用。在优化的测试条件下,该免疫传感器在 1 fg mL 至 1 μg mL 的范围内呈现出良好的线性关系,检测限为 0.8 fg mL(相当于 10 μL 样品中约 134 个分子)。基于 eATRP 信号放大技术的 ECL 免疫传感系统为通过检测 CYFRA 21-1 快速诊断肺癌提供了一种新方法。本文通过 eATRP 信号放大策略制备了用于超灵敏检测 CYFRA 21-1 的电化学发光生物传感器,该传感器具有灵敏度高、重现性好等优点,在分析低丰度方面具有潜在的前景。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/864d/7940335/549cf25b44d0/604_2020_4677_Figa_HTML.jpg

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