Liang Ke-Zhong, Qi Jun-Sheng, Mu Wei-Jun, Chen Zai-Gang
Department of Chemistry and Environment, Chongqing Three Gorges University, Chongqing 400400, PR China.
J Biochem Biophys Methods. 2008 Apr 24;70(6):1156-62. doi: 10.1016/j.jprot.2007.11.007. Epub 2007 Nov 24.
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 3delta). 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.
一种基于将睾酮抗体包封到掺杂金纳米线的聚乙烯醇缩丁醛溶胶 - 凝胶膜中的直接、快速且无标记的睾酮电化学免疫分析方法已被报道。使用纳米孔聚碳酸酯膜制备的金纳米线用于将睾酮抗体缀合到探针表面。金纳米线的存在为生物分子提供了生物相容性微环境，极大地放大了电极表面生物分子的固定量，并提高了免疫传感器的灵敏度。与金纳米颗粒缀合探针相比，金纳米线功能化探针可避免生物分子从复合膜中泄漏，并增强了传感器的稳定性。详细研究了所得免疫传感器的性能及影响其性能的因素。在最佳条件下，所开发的免疫传感器与睾酮在1.2至83.5 ng mL(-1)范围内呈现良好的线性关系，检测限为0.1 ng mL(-1)（3δ）。此外，所提出的免疫传感器具有高灵敏度、良好的重现性和长期稳定性。所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，所开发的免疫分析方法在临床诊断中检测睾酮方面具有有前景的替代方法。与传统酶联免疫吸附测定法相比，所提出的免疫分析方法简单快速，无需多个标记和分离步骤。重要的是，该途径为将金纳米线纳入固体基质用于生物传感应用提供了一种替代方法。
