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基于聚苯胺保护的金纳米粒子的媒介物和无标记电化学皮质醇生物传感器。

Polyaniline protected gold nanoparticles based mediator and label free electrochemical cortisol biosensor.

机构信息

Bio-MEMS and Microsystem Lab, Department of Electrical Engineering, University of South Florida, Tampa, FL 33620, United States.

出版信息

Biosens Bioelectron. 2011 Oct 15;28(1):166-73. doi: 10.1016/j.bios.2011.07.015. Epub 2011 Jul 20.

Abstract

Polyaniline protected gold nanoparticles (PPAuNPs) were electrophoretically deposited onto a gold electrode, and utilized to fabricate an electrochemical cortisol biosensor. Cortisol specific monoclonal antibody (C-Mab) was covalently immobilized onto the surface of a PPAuNP/Au electrode using N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide (EDC/NHS) chemistry. BSA was employed for blocking nonspecific adsorption on the electrode surface. PPAuNP formation and BSA/C-Mab/PPAuNP/Au electrode fabrication were characterized using transmission electron microscopy, atomic force microscopy and electrochemical impedance techniques, respectively. Cyclic voltammetry and differential pulse voltammetric techniques were used to determine the cortisol concentration in a phosphate buffer saline (PBS) solution. Results confirmed that the PPAuNP based electrode was stable during repeated scans and exhibited repeatable redox peaks. Further, the BSA/C-Mab/PPAuNP/Au electrode in the PBS buffer accurately detected cortisol in the range of 1 pM-100 nM with a sensitivity of 1.63 μAM(-1). The biosensor was found to be selective against BSA and 17-α-hydroxy progesterone. This research establishes the feasibility of using a PPAuNP based matrix for a label and mediator free electrochemical biosensor for cortisol, a stress biomarker.

摘要

聚苯胺保护的金纳米粒子(PPAuNP)通过电泳沉积在金电极上,并用于构建电化学皮质醇生物传感器。皮质醇特异性单克隆抗体(C-Mab)通过 N-乙基-N'-(3-二甲基氨基丙基)碳二亚胺和 N-羟基琥珀酰亚胺(EDC/NHS)化学被共价固定在 PPAuNP/Au 电极表面。BSA 用于阻止电极表面的非特异性吸附。PPAuNP 的形成和 BSA/C-Mab/PPAuNP/Au 电极的制备分别通过透射电子显微镜、原子力显微镜和电化学阻抗技术进行了表征。循环伏安法和差分脉冲伏安法用于在磷酸盐缓冲盐水(PBS)溶液中测定皮质醇浓度。结果证实,基于 PPAuNP 的电极在重复扫描过程中稳定,并且表现出可重复的氧化还原峰。此外,BSA/C-Mab/PPAuNP/Au 电极在 PBS 缓冲液中能够准确检测 1 pM-100 nM 范围内的皮质醇,灵敏度为 1.63 μAM(-1)。该生物传感器对 BSA 和 17-α-羟基孕酮具有选择性。这项研究确立了使用基于 PPAuNP 的基质作为无标记和无介体的电化学生物传感器来检测皮质醇(一种应激生物标志物)的可行性。

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