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基于靶标诱导光致猝灭的溶菌酶简便电化学生物传感器

A facile electrochemical aptasensor for lysozyme detection based on target-induced turn-off of photosensitization.

机构信息

College of Materials and Chemistry & Chemical Engineering, Chengdu University of Technology, Chengdu 610059, China.

College of Environment and Ecology, Chengdu University of Technology, Chengdu 610059, China; State Environmental Protection Key Laboratory of Synergetic Control and Joint Remediation for Soil & Water Pollution, Chengdu University of Technology, Chengdu 610059, China.

出版信息

Biosens Bioelectron. 2019 Feb 1;126:412-417. doi: 10.1016/j.bios.2018.09.074. Epub 2018 Sep 21.

Abstract

The quantification of proteins is essential in fundamental research or clinical applications. Here, we developed a facile electrochemical aptasensor based on target-induced turn-off of photosensitization for label-free and ultrasensitive detection of protein (exemplified by lysozyme). EB (ethidium bromide) molecules that were embedded in dsDNA between lysozyme binding aptamer and complementary DNA immobilized on the electrode, could photo-cleave the dsDNA via singlet oxygen (O) during photosensitization, resulting in a high voltammetry current of the [Fe(CN)]. Upon recognition of the lysozyme by aptamer, the EB molecules were released from dsDNA, and its photosensitization activity was turned off. As a result, more amount of complementary DNA was retained on the Au nanoparticles modified carbon nanotube paste electrode (AuNPs-CNPE), leading to a declined voltammetry current. Such a sensing strategy allowed detection of 10 pM-1 µM lysozyme with a low detection limit (about 2 pM). Besides, the sensor was free of labeling procedure as well as extra signal amplification step, and the CNPE modification was quite simple, only with AuNPs. The sensor also showed excellent selectivity toward lysozyme in the presence of interfering proteins, such as thrombin, bovine serum albumin, myoglobin, etc. The proposed sensor was applied to the determination of lysozyme in urine samples with the recoveries ranging from 96.6% to 101%. The proposed biosensor holds a great promise in developing other electrochemical sensors based on photosensitization.

摘要

蛋白质的定量分析在基础研究或临床应用中至关重要。在这里,我们开发了一种基于目标诱导光致猝灭的简便电化学适体传感器,用于无标记和超灵敏检测蛋白质(以溶菌酶为例)。嵌入在溶菌酶结合适体和固定在电极上的互补 DNA 之间的 dsDNA 中的 EB(溴化乙锭)分子可以在光致猝灭过程中通过单线态氧(O)切割 dsDNA,导致 [Fe(CN)。当适体识别溶菌酶时,EB 分子从 dsDNA 中释放出来,其光致猝灭活性被关闭。结果,更多数量的互补 DNA 保留在修饰有金纳米粒子的碳纳米管糊电极(AuNPs-CNPE)上,导致伏安电流下降。这种传感策略允许检测 10 pM-1 μM 的溶菌酶,检测限低至约 2 pM。此外,传感器无需标记程序和额外的信号放大步骤,并且 CNPE 修饰非常简单,仅使用 AuNPs。该传感器在存在干扰蛋白(如凝血酶、牛血清白蛋白、肌红蛋白等)时对溶菌酶也表现出优异的选择性。该传感器已应用于尿液样品中溶菌酶的测定,回收率在 96.6%至 101%之间。该传感器有望在开发基于光致猝灭的其他电化学传感器方面取得进展。

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