Department of Analytical Chemistry, Faculty of Chemistry, University Complutense of Madrid, 28040 Madrid, Spain.
Talanta. 2014 Jan;118:61-7. doi: 10.1016/j.talanta.2013.10.008. Epub 2013 Oct 9.
A novel electrochemical immunosensor for the determination of ceruloplasmin (Cp) in human serum and urine is reported. The immunosensor configuration involves an indirect competitive immunoassay implying covalent immobilization of Cp on activated carboxylic groups at carbon nanotubes-modified screen-printed electrodes (CNTs/SPE). After Cp immobilization and reaction between the target analyte and anti-ceruloplasmin antibodies in solution, the remaining non-conjugated antibody is attached on the Cp-CNTs modified electrode. Monitoring of Cp is performed by means of a secondary antibody labeled with peroxidase (HRP-anti-IgG) and measurement of the amperometric current resulting from the addition of hydrogen peroxide in the presence of hydroquinone as the redox mediator. The experimental variables affecting the analytical performance of the immunosensor were optimized. Calibration curves for Cp provided a linear range between 0.07 and 250 μg/mL (r=0.997). The limit of detection achieved was 21 ng/mL. These analytical characteristics allow the immunosensor to be successfully used for the determination of Cp in spiked human serum and urine at various concentration levels.
本文报道了一种用于测定人血清和尿液中铜蓝蛋白（Cp）的新型电化学免疫传感器。该免疫传感器的配置涉及一种间接竞争免疫分析，该分析涉及将 Cp 共价固定在经碳纳米管修饰的丝网印刷电极（CNTs/SPE）上的活化羧酸基团上。在 Cp 固定化以及目标分析物与溶液中抗铜蓝蛋白抗体之间的反应之后，剩余的未缀合抗体附着在 Cp-CNTs 修饰电极上。通过用过氧化物酶标记的二次抗体（HRP-抗-IgG）进行监测，并在存在氢醌作为氧化还原介体的情况下添加过氧化氢来测量安培电流，从而实现对 Cp 的监测。优化了影响免疫传感器分析性能的实验变量。Cp 的校准曲线在 0.07 和 250 μg/mL 之间呈线性关系（r=0.997）。检测限达到 21 ng/mL。这些分析特性允许免疫传感器成功用于测定各种浓度水平的人血清和尿液中的 Cp。
