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基于适配体功能化石墨烯修饰 SPR 界面的溶菌酶检测。

Lysozyme detection on aptamer functionalized graphene-coated SPR interfaces.

机构信息

Institut de Recherche Interdisciplinaire (IRI), CNRS USR 3078, Université Lille1, Parc de la Haute Borne, 50 avenue de Halley, BP 70478, 59658 Villeneuve d'Ascq, France.

出版信息

Biosens Bioelectron. 2013 Dec 15;50:239-43. doi: 10.1016/j.bios.2013.06.026. Epub 2013 Jun 27.

Abstract

The paper reports on a surface plasmon resonance (SPR)-based approach for the sensitive and selective detection of lysozyme. The SPR sensor consists of a 50 nm gold film coated with a thin film of reduced graphene oxide (rGO) functionalized with anti-lysozyme DNA aptamer. The SPR chip coating with rGO matrix was achieved through electrophoretic deposition of graphene oxide (GO) at 150 V. Electrophoretic deposition resulted in partial reduction of GO to rGO with a thickness depending on the deposition time. For very short time pulses of 20 s, the resulting rGO film had a thickness of several nanometers and was appropriate for SPR sensing. The utility of the graphene-based SPR sensor for the selective and sensitive detection of proteins was demonstrated using lysozyme as model protein. Functionalization of rGO matrix with anti-lysozyme DNA aptamer through π-stacking interactions allowed selective SPR detection of lysozyme. The graphene-based SPR biosensor provides a means for the label-free, concentration-dependent and selective detection of lysozymes with a detection limit of 0.5 nM.

摘要

本文报道了一种基于表面等离子体共振(SPR)的方法,用于灵敏和选择性检测溶菌酶。SPR 传感器由 50nm 厚的金膜组成,表面涂有一层经溶菌酶 DNA 适体功能化的还原氧化石墨烯(rGO)薄膜。通过在 150V 下电泳沉积氧化石墨烯(GO)实现了带有 rGO 基质的 SPR 芯片涂覆。电泳沉积导致 GO 部分还原为 rGO,其厚度取决于沉积时间。对于 20 秒的非常短的脉冲时间,得到的 rGO 薄膜厚度为几纳米,适用于 SPR 感测。使用溶菌酶作为模型蛋白,证明了基于石墨烯的 SPR 传感器在选择性和灵敏检测蛋白质方面的应用。通过π-堆积相互作用将 rGO 基质功能化,与抗溶菌酶 DNA 适体结合,实现了对溶菌酶的选择性 SPR 检测。基于石墨烯的 SPR 生物传感器提供了一种无需标记、浓度依赖性和选择性检测溶菌酶的方法,检测限为 0.5nM。

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