†CAS Key Laboratory of Soft Matter Chemistry, Collaborative Innovation Center of Chemistry for Energy Materials, Department of Chemistry, University of Science and Technology of China, Hefei, Anhui 230026, P. R. China.
‡Zhengzhou Institute of Multipurpose Utilization of Mineral Resources, Chinese Academy of Geological Sciences, Zhengzhou 450006, P. R. China.
ACS Appl Mater Interfaces. 2015 Apr 15;7(14):7599-604. doi: 10.1021/am509094p. Epub 2015 Mar 31.
A novel electrochemiluminescence (ECL) immunosensor was developed for the determination of N-terminal pro-brain natriuretic peptide (NT-proBNP) by using N-(aminobutyl)-N-(ethylisoluminol) (ABEI)-functionalized gold nanodots/chitosan/multiwalled carbon nanotubes (ABEI/GNDs/chitosan/COOH-MWCNTs) hybrid as nanointerface. First, ABEI/GNDs/chitosan/COOH-MWCNTs hybrid nanomaterials were grafted onto the surface of ITO electrode via the film-forming property of hybrid nanomaterials. The anti-NT-proBNP antibody was connected to the surface of modified electrode by virtue of amide reaction via glutaraldehyde. The obtained sensing platform showed strong and stable ECL signal. When NT-proBNP was captured by its antibody immobilized on the sensing platform via immunoreaction, the ECL intensity decreased. Direct ECL signal changes were used for the determination of NT-proBNP. The present ECL immunosensor demonstrated a quite wide linear range of 0.01-100 pg/mL. The achieved low detection limit of 3.86 fg/mL was about 3 orders of magnitude lower than that obtained with electrochemistry method reported previously. Because of the simple and fast analysis, high sensitivity and selectivity, and stable and reliable response, the present immunosensor has been successfully applied to quantify NT-proBNP in practical plasma samples. The success of the sensor in this work also confirms that ABEI/GNDs/chitosan/COOH-MWCNTs hybrid is an ideal nanointerface to fabricate a sensing platform. Furthermore, the proposed strategy could be applied in the detection of other clinically important biomarkers.
一种新型的电化学发光(ECL)免疫传感器,通过使用 N-(氨丁基)-N-(乙基异鲁米诺)(ABEI)功能化金纳米点/壳聚糖/多壁碳纳米管(ABEI/GNDs/壳聚糖/COOH-MWCNTs)杂化作为纳米界面来测定 N-末端脑利钠肽前体(NT-proBNP)。首先,通过杂化纳米材料的成膜性质将 ABEI/GNDs/壳聚糖/COOH-MWCNTs 杂化纳米材料接枝到 ITO 电极表面。通过戊二醛的酰胺反应将抗 NT-proBNP 抗体连接到修饰电极的表面。所得的传感平台表现出强而稳定的 ECL 信号。当 NT-proBNP 通过免疫反应被固定在传感平台上的抗体捕获时,ECL 强度降低。直接 ECL 信号变化用于测定 NT-proBNP。该 ECL 免疫传感器表现出相当宽的线性范围为 0.01-100 pg/mL。实现的低检测限为 3.86 fg/mL,比以前报道的电化学方法低约 3 个数量级。由于分析简单快速、灵敏度和选择性高、响应稳定可靠,该免疫传感器已成功应用于实际血浆样品中 NT-proBNP 的定量。该传感器在这项工作中的成功也证实了 ABEI/GNDs/壳聚糖/COOH-MWCNTs 杂化是构建传感平台的理想纳米界面。此外,所提出的策略可应用于其他临床重要生物标志物的检测。