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基于多巴胺功能化 CuInS/ZnS 量子点的酪氨酸酶诱导荧光免疫分析用于检测 tau 蛋白

A tyrosinase-induced fluorescence immunoassay for detection of tau protein using dopamine-functionalized CuInS/ZnS quantum dots.

机构信息

School of Pharmacy, Fujian Medical University, Fuzhou, 350122, Fujian, China.

School of Basic Medical Sciences, Fujian Medical University, Fuzhou, 350122, Fujian, China.

出版信息

Anal Bioanal Chem. 2019 Aug;411(20):5277-5285. doi: 10.1007/s00216-019-01909-9. Epub 2019 Jun 3.

Abstract

Rapid, highly sensitive detection of tau protein and other neurodegenerative biomarkers remains a significant hurdle for diagnostic tests for Alzheimer's disease. In this work, we developed a novel tyrosinase (TYR)-induced tau aptamer-tau-tau antibody (anti-tau) sandwich fluorescence immunoassay to detect tau protein that used dopamine (DA)-functionalized CuInS/ZnS quantum dots as the fluorophore. CuInS/ZnS core/shell quantum dots with high luminescence, low toxicity, and excellent biocompatibility were successfully fabricated and decorated with DA through amide conjugation. Meanwhile, TYR was conjugated with anti-tau by a click reaction. When DA-functionalized CuInS/ZnS quantum dots were added to the sandwich system, TYR catalyzed the transformation of DA to dopamine quinone, which acted as an effective electron acceptor and triggered fluorescence quenching. The fluorescence intensity of the immunoassay based on DA-functionalized CuInS/ZnS quantum dots shows good performance in terms of linearity with the logarithm of tau protein concentration, with a linear concentration range from 10 pM to 200 nM. This work is the first to use a TYR-induced fluorescence immunoassay for the rapid detection of tau protein, paving a new way for the detection of disease biomarkers. Graphical abstract.

摘要

快速、高灵敏度地检测 tau 蛋白和其他神经退行性生物标志物仍然是阿尔茨海默病诊断测试的一个重大障碍。在这项工作中,我们开发了一种新型的酪氨酸酶(TYR)诱导的 tau 适体- tau- tau 抗体(抗 tau)夹心荧光免疫分析方法,用于检测 tau 蛋白,该方法使用多巴胺(DA)功能化的 CuInS/ZnS 量子点作为荧光团。成功制备了具有高发光、低毒性和优异生物相容性的 CuInS/ZnS 核/壳量子点,并通过酰胺偶联将 DA 修饰到其上。同时,通过点击反应将 TYR 与抗 tau 偶联。当将 DA 功能化的 CuInS/ZnS 量子点添加到夹心体系中时,TYR 催化 DA 向多巴胺醌的转化,多巴胺醌作为有效的电子受体触发荧光猝灭。基于 DA 功能化的 CuInS/ZnS 量子点的免疫分析的荧光强度在 tau 蛋白浓度的对数线性范围内表现出良好的性能,线性浓度范围为 10 pM 至 200 nM。这项工作首次使用 TYR 诱导的荧光免疫分析来快速检测 tau 蛋白,为疾病生物标志物的检测开辟了新途径。

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