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基于能量转移的氧化石墨烯平台用于蛋白酶活性检测。

A graphene oxide platform for energy transfer-based detection of protease activity.

机构信息

The Key Lab of Analysis and Detection Technology for Food Safety of the MOE, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, College of Chemistry and Chemical Engineering, Fuzhou University, Fuzhou 350002, PR China.

出版信息

Biosens Bioelectron. 2011 May 15;26(9):3894-9. doi: 10.1016/j.bios.2011.03.003. Epub 2011 Mar 8.

Abstract

In this article, we report the first graphene oxide (GO)-based platform to detect protease activity in a homogeneous real-time format. In designing such GO-based biosensing platform, we put a protease substrate peptide as the linker between the energy transfer donor (QDs) and the energy transfer acceptor (GO) to fabricate the GO-peptide-QDs nanoprobes. In the nanoprobes, the photoluminescence (PL) of donor QDs was strongly quenched due to the presence of GO in close proximity. The protease activity caused modulation in the efficiency of the energy transfer between the acceptor and donor, thus enabling the protease assay. The proposed GO-based platform is easy to assemble and has little background interference, yet still give superior sensitivity and rapid response. Furthermore, this GO-QDs architecture can serve as a universal platform by simply changing the types of peptide sequences for the different proteases. In this work, GO-based platform has been successfully applied in the sensitive detection of matrix metalloproteinase (MMP) and thrombin activity. Meanwhile, we also utilized this platform to monitor the protease inhibitor. The proposed GO-based platform is anticipated to find applications in the diagnosis of protease-related diseases and screening of potential drugs with high sensitivity in a high-throughput way.

摘要

在本文中,我们报告了第一个基于氧化石墨烯(GO)的平台,用于以均相实时格式检测蛋白酶活性。在设计这种基于 GO 的生物传感平台时,我们将蛋白酶底物肽用作能量转移供体(QDs)和能量转移受体(GO)之间的连接体,以制备 GO-肽-QDs 纳米探针。在纳米探针中,由于存在近距离的 GO,供体 QDs 的光致发光(PL)被强烈猝灭。蛋白酶活性导致受体和供体之间能量转移效率的调制,从而能够进行蛋白酶测定。所提出的基于 GO 的平台易于组装,背景干扰小,但仍具有优异的灵敏度和快速响应。此外,通过简单地改变肽序列的类型,这种 GO-QDs 结构可以作为通用平台用于不同的蛋白酶。在这项工作中,基于 GO 的平台已成功应用于基质金属蛋白酶(MMP)和凝血酶活性的灵敏检测。同时,我们还利用该平台监测蛋白酶抑制剂。预计该基于 GO 的平台将在蛋白酶相关疾病的诊断和高通量筛选潜在药物方面得到应用,具有高灵敏度。

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