生物分子/金纳米线掺杂溶胶-凝胶膜用于睾酮的无标记电化学免疫分析

Ke-Zhong Liang , Jun-Sheng Qi , Wei-Jun Mu , Zai-Gang Chen
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引用次数: 54

摘要

本文描述了一种直接、快速、无标记的睾酮电化学免疫分析方法,该方法基于将睾酮抗体包封在掺杂金纳米线的聚乙烯醇丁醛溶胶-凝胶膜中。用纳米孔聚碳酸酯膜制备的金纳米线将睾酮抗体偶联到探针表面。金纳米线的存在为生物分子提供了生物相容性微环境,极大地增加了生物分子在电极表面的固定化量,提高了免疫传感器的灵敏度。与金纳米粒子偶联探针相比,金纳米线功能化探针可以避免生物分子从复合膜中泄漏,增强传感器的稳定性。详细研究了该免疫传感器的性能及其影响因素。在最佳条件下,所开发的免疫传感器与睾酮表现出良好的线性关系,检测限为0.1 ng mL - 1(在3δ),范围为1.2 ~ 83.5 ng mL - 1。该免疫传感器具有灵敏度高、重复性好、长期稳定等特点。制备的免疫传感器用于分析人血清标本中的睾酮。分析结果表明,开发的免疫分析法有一个很有前途的替代方法检测睾酮在临床诊断。与传统elisa相比,该方法简便、快速,无需多次标记和分离步骤。重要的是,该路线提供了一种将金纳米线纳入生物传感应用的固体基质的替代方法。
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Biomolecules/gold nanowires-doped sol–gel film for label-free electrochemical immunoassay of testosterone

A direct, rapid, and label-free electrochemical immunoassay method for testosterone has been described based on encapsulating testosterone antibody into polyvinyl butyral sol–gel film doped with gold nanowires. Gold nanowires prepared by using nanopore polycarbonate membrane were used to conjugate testosterone antibody onto the probe surface. The presence of gold nanowires provided a biocompatible microenvironment for biomolecules, greatly amplified the immobilized amount of biomolecules on the electrode surface, and improved the sensitivity of the immunosensor. In comparison with gold nanoparticle-conjugating probe, the gold nanowire-functionalized probe could avoid the leakage of biomolecules from the composite film, and enhanced the stability of the sensor. The performance and factors influencing the performance of the resulting immunosensor were investigated in detail. Under optimal conditions, the developed immunosensor exhibited a good linear relationship with testosterone ranging from 1.2 to 83.5 ng mL 1 with a detection limit of 0.1 ng mL 1 (at 3δ). Moreover, the proposed immunosensor exhibited high sensitivity, good reproducibility and long-term stability. The as-prepared immunosensors were used to analyze testosterone in human serum specimens. Analytical results suggest that the developed immunoassay has a promising alternative approach for detecting testosterone in the clinical diagnosis. Compared with the conventional ELISAs, the proposed immunoassay method was simple and rapid without multiple labeling and separation steps. Importantly, the route provides an alternative approach to incorporate gold nanowires into the solid matrix for biosensing application.

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