Department of Analytical Chemistry, Faculty of Chemistry, University Complutense of Madrid, Spain.
Department of Analytical Chemistry, Faculty of Chemistry, University Complutense of Madrid, Spain.
Bioelectrochemistry. 2020 Jun;133:107484. doi: 10.1016/j.bioelechem.2020.107484. Epub 2020 Feb 14.
Electro-click methodology was employed to prepare an electrochemical immunosensor for the cytokine interleukin 1β (IL-1β). The strategy involved binding of ethynylated IgG to azide-MWCNTs modified electrodes by Cu(I) catalyzed-cycloaddition reaction where the catalyst was electrochemically synthesized. This electro-click protocol is significantly faster and greener than the methods for catalyst generation through chemical reduction. The oriented immobilization of the capture antibody onto IgG-MWCNTs conjugates allowed the preparation of a sandwich-type immunosensor using biotinylated anti-IL-1β as detector antibody labeled with alkaline phosphatase-streptavidin (AP-strept). Differential pulse voltammetric transduction through the 1-naphthylphosphate/1-naphthol system was carried out. The analytical characteristics achieved with the electrochemical immunosensor showed a calibration curve exhibiting two linear ranges between 10 and 200 pg mL (r = 0.998), and from 200 to 1200 pg mL (r = 0.998), and a LOD value of 5.2 pg mL, an improvement compared with those claimed for commercial ELISA kits. In addition, the assay time was at least one hour shorter. Excellent performance was observed in the determination of IL-1β in saliva with no need for sample treatment, and by simple interpolation using a calibration plot constructed with standard solutions of the target cytokine.
电化学点击法被用于制备用于细胞因子白细胞介素 1β（IL-1β）的电化学免疫传感器。该策略涉及通过 Cu（I）催化的环加成反应将乙炔基化 IgG 结合到叠氮化物-MWCNTs 修饰的电极上，其中催化剂通过电化学合成。与通过化学还原生成催化剂的方法相比，这种电点击方案明显更快且更环保。通过将捕获抗体定向固定在 IgG-MWCNTs 缀合物上，使用生物素化抗 IL-1β作为碱性磷酸酶-链霉亲和素（AP-链霉）标记的检测抗体，可以制备夹心型免疫传感器。通过 1-萘基磷酸/1-萘酚体系进行差分脉冲伏安法检测。电化学免疫传感器的分析特性显示出两条线性范围在 10 至 200 pg mL（r = 0.998）和 200 至 1200 pg mL（r = 0.998）之间的校准曲线，以及 5.2 pg mL 的检测限，与商业 ELISA 试剂盒声称的相比有所提高。此外，测定时间至少缩短了一个小时。在唾液中测定 IL-1β 时，该测定方法表现出优异的性能，无需进行样品处理，并且通过使用目标细胞因子的标准溶液构建校准曲线进行简单插值即可实现。
