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A bimetallic PtPd hybrid nanostructure-amplified enzyme-free conductometric immunoassay for lipocalin-2 in renal cell carcinoma on an interdigitated micro-comb electrode
Analytical Methods ( IF 2.7 ) Pub Date : 2020-03-19 , DOI: 10.1039/c9ay02525a Chaoqun Huang 1, 2, 3, 4, 5 , Fengling Zhang 1, 2, 3, 4, 5 , Qingshui Wang 5, 6, 7, 8 , Yao Lin 5, 6, 7, 8 , Jiyi Huang 1, 2, 3, 4, 5
Analytical Methods ( IF 2.7 ) Pub Date : 2020-03-19 , DOI: 10.1039/c9ay02525a Chaoqun Huang 1, 2, 3, 4, 5 , Fengling Zhang 1, 2, 3, 4, 5 , Qingshui Wang 5, 6, 7, 8 , Yao Lin 5, 6, 7, 8 , Jiyi Huang 1, 2, 3, 4, 5
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A new enzyme-free conductometric immunoassay based on bimetallic PtPd hybrid nanostructures was developed for the sensitive determination of lipocalin-2 in renal cell carcinoma. An immunosensor was prepared by immobilizing the capture antibody on an interdigitated micro-comb electrode, whereas bimetallic PtPd hybrid nanostructures were utilized for the conjugation of the detection antibody. Initially, a sandwich immunoreaction was carried out between the capture antibody and detection antibody in the presence of lipocalin-2 on the functional interdigitated micro-comb electrode. Thereafter, the immobilized PtPd bimetallic nanostructures could be used as enzymatic mimics to catalyze the reaction of hydrogen peroxide (H2O2) in 0.01 M phosphate buffer solution (pH 7.4) containing KI, thereby resulting in the local variation in the conductivity of the modified electrode. Several labeling strategies using Pt nanoparticles, Pd nanoparticles and bimetallic PtPd hybrid nanostructures were investigated for the detection of lipocalin-2, and improved analytical features were acquired with PtPd bimetallic nanostructures. With the PtPd labeling method, the factors influencing the analytical performance of the conductometric immunoassay were studied in detail. The strong attachment of biomolecules to the interdigitated micro-comb electrode and PtPd bimetallic nanostructures enabled a good repeatability and intermediate precision down to 10.4%. The dynamic concentration range spanned from 10 pg mL−1 to 30 ng mL−1 with a low detection limit of 5.9 pg mL−1 lipocalin-2 under optimum conditions. The reliability of the conductometric immunoassay was verified for the detection of lipocalin-2 in 4 blood serum and 4 urine samples and matched well with that obtained from the commercial lipocalin-2 enzyme-linked immunosorbant assay kit.
中文翻译：
双金属PtPd杂合纳米结构扩增的无酶电导免疫测定法在指状微梳电极上检测肾细胞癌中的lipocalin-2
建立了一种基于双金属PtPd杂化纳米结构的新型无酶电导免疫测定方法，用于灵敏测定肾癌中lipocalin-2。通过将捕获抗体固定在指状微梳状电极上来制备免疫传感器，而将双金属PtPd杂化纳米结构用于检测抗体的偶联。最初，在脂叉蛋白2的存在下，在功能交叉指状微梳状电极上，在捕获抗体和检测抗体之间进行了夹心免疫反应。此后，固定化的PtPd双金属纳米结构可以用作酶模拟物，以催化过氧化氢（H 2 O 2）在含有KI的0.01 M磷酸盐缓冲溶液（pH 7.4）中，从而导致修饰电极的电导率局部变化。研究了使用Pt纳米颗粒，Pd纳米颗粒和双金属PtPd杂化纳米结构的几种标记策略，用于lipocalin-2的检测，并利用PtPd双金属纳米结构获得了改进的分析特性。使用PtPd标记方法，详细研究了影响电导免疫测定分析性能的因素。生物分子与指状微梳状电极和PtPd双金属纳米结构的牢固结合使得其具有良好的重复性和低至10.4％的中间精度。动态浓度范围从10 pg mL -1到30 ng mL -1在最佳条件下的最低检测限为5.9 pg mL -1 lipocalin-2。验证了电导免疫测定法在4个血清和4个尿液样品中检测lipocalin-2的可靠性，并与从商业lipocalin-2酶联免疫吸附测定试剂盒中获得的脂溶性蛋白非常匹配。
更新日期：2020-04-24
中文翻译：
双金属PtPd杂合纳米结构扩增的无酶电导免疫测定法在指状微梳电极上检测肾细胞癌中的lipocalin-2
建立了一种基于双金属PtPd杂化纳米结构的新型无酶电导免疫测定方法，用于灵敏测定肾癌中lipocalin-2。通过将捕获抗体固定在指状微梳状电极上来制备免疫传感器，而将双金属PtPd杂化纳米结构用于检测抗体的偶联。最初，在脂叉蛋白2的存在下，在功能交叉指状微梳状电极上，在捕获抗体和检测抗体之间进行了夹心免疫反应。此后，固定化的PtPd双金属纳米结构可以用作酶模拟物，以催化过氧化氢（H 2 O 2）在含有KI的0.01 M磷酸盐缓冲溶液（pH 7.4）中，从而导致修饰电极的电导率局部变化。研究了使用Pt纳米颗粒，Pd纳米颗粒和双金属PtPd杂化纳米结构的几种标记策略，用于lipocalin-2的检测，并利用PtPd双金属纳米结构获得了改进的分析特性。使用PtPd标记方法，详细研究了影响电导免疫测定分析性能的因素。生物分子与指状微梳状电极和PtPd双金属纳米结构的牢固结合使得其具有良好的重复性和低至10.4％的中间精度。动态浓度范围从10 pg mL -1到30 ng mL -1在最佳条件下的最低检测限为5.9 pg mL -1 lipocalin-2。验证了电导免疫测定法在4个血清和4个尿液样品中检测lipocalin-2的可靠性，并与从商业lipocalin-2酶联免疫吸附测定试剂盒中获得的脂溶性蛋白非常匹配。
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