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抗体在仿生结构上的直接固定作为传感平台。

Direct immobilisation of antibodies on a bioinspired architecture as a sensing platform.

机构信息

Key Lab of Corrosion Science, Shandong Province, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, China.

出版信息

Biosens Bioelectron. 2011 Jan 15;26(5):2595-600. doi: 10.1016/j.bios.2010.11.013. Epub 2010 Nov 26.

Abstract

A sensitive and selective immunosensor for the nonlabeled detection of sulfate-reducing bacteria (SRB) is constructed using a self-polymerised polydopamine film as the immobilisation platform. Self-polymerisation of dopamine is used as a powerful approach for applying multifunctional coatings onto the surface of a gold electrode. The polydopamine film is used not only as the immobilisation platform, but also as a cross-linker reagent for the immobilisation of the anti-SRB antibody. The polydopamine film is loaded with a high density of anti-SRB antibodies linked to the substrate to obtain high response signals. The formation and fabrication of the biosensor and the quantification of antibody anchoring are monitored, and SRB detection is performed by either quartz crystal microbalance (QCM) or electrochemical impedance spectroscopy (EIS). After modeling the impedance Nyquist plots of the SRB/anti-SRB/polydopamine/gold electrode for increasing concentrations of SRB, the electron transfer resistance (R(ct)) is used as a measure of immunocomplex binding. The R(ct) is correlated with the concentration of bacterial cells in the range of 1.8×10(2) to 1.8×10(6) CFU mL(-1); the detection limit is 50 CFU mL(-1). This work demonstrates a new immobilisation platform for the development of a sensitive and label-less impedimetric and piezoelectric immunosensor. This immunosensor may be broadly applied in clinical diagnoses and the monitoring of water environmental pollution. The method proposed is distinct in its ease of application, use of a simple protocol, and mild reaction conditions. These allow it to be applied to a wide variety of materials.

摘要

一种基于自聚合多巴胺膜作为固定化平台的灵敏、选择性免疫传感器,用于非标记检测硫酸盐还原菌(SRB)。多巴胺的自聚合被用作在金电极表面施加多功能涂层的有力方法。聚多巴胺膜不仅用作固定化平台,还用作固定化抗 SRB 抗体的交联试剂。聚多巴胺膜负载有高密度与基底相连的抗 SRB 抗体,以获得高响应信号。监测生物传感器的形成和制造以及抗体锚定的定量,并通过石英晶体微天平(QCM)或电化学阻抗谱(EIS)进行 SRB 检测。在对 SRB/抗 SRB/聚多巴胺/金电极的阻抗奈奎斯特图进行建模以增加 SRB 的浓度后,电子转移电阻(R(ct))用作免疫复合物结合的测量。R(ct)与细菌细胞浓度在 1.8×10(2)到 1.8×10(6) CFU mL(-1)范围内相关;检测限为 50 CFU mL(-1)。这项工作展示了一种用于开发灵敏、无标记阻抗和压电免疫传感器的新型固定化平台。这种免疫传感器可广泛应用于临床诊断和水环境污染监测。所提出的方法在其易于应用、使用简单的方案和温和的反应条件方面具有独特性。这些允许它应用于各种材料。

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