Key Laboratory of Chemical Sensing & Analysis in Universities of Shandong (University of Jinan), School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022, China.
Biosens Bioelectron. 2013 Nov 15;49:111-7. doi: 10.1016/j.bios.2013.05.010. Epub 2013 May 13.
A simple, low-cost and sensitive origami electrochemical immunoassay-device was developed based on a novel gold nanoparticle modified porous paper working electrode (Au-PWE) for point-of-care testing. Azide-functionalized Au-PWE was prepared by the functionalization of Au-PWE with 1-azidoundecan-11-thiol. Alkyne end-terminated antibody was prepared with 4-pentynoic acid and antibody by the 1-ethyl-3-(3-(dimethylamino) propyl) carbodiimide hydrochloride and N-hydroxysuccinimide activation reaction. Alkyne-antibody was coupled to azido-Au-PWE by click reaction as a recognition element. Nearly monodispersed sphere-like silica-coated ferroferric oxide (Fe3O4@SiO2) nanoparticles were prepared via the reverse microemulsion method. Azide-functionalized Fe3O4@SiO2 was prepared by the functionalization of silica shell with 3-bromopropyltrichlorosilane followed by substitution with sodium azide. Alkyne-functionalized antibody and horse radish peroxidase were coupled to azide-functionalized Fe3O4@SiO2 by click reaction as signal label. Horse radish peroxidase and ferroferric oxide could catalyze the oxidation of thionine in the presence of hydrogen peroxide. After the sandwich immunoreaction, the current was proportional to the logarithm of the Microcystin-LR. The linear regression equation was i(μA)=119.89+46.27 log cMC-LR (μg/mL) in the range from 0.01 to 200 μg/mL. The limit of detection was 0.004 μg/mL. This immunoassay would provide a universal immunoassay method in environmental monitoring and public health.
一种基于新型金纳米粒子修饰的多孔纸工作电极（Au-PWE）的简单、低成本和灵敏的折纸电化学生物免疫分析装置，用于即时检测。通过 1-叠氮十一烷-11-硫醇对 Au-PWE 进行功能化，制备叠氮功能化的 Au-PWE。通过 1-乙基-3-(3-(二甲基氨基)丙基)碳化二亚胺盐酸盐和 N-羟基琥珀酰亚胺的活化反应，用 4-戊炔酸和抗体制备炔基末端抗体。炔基抗体通过点击反应与叠氮 Au-PWE 偶联作为识别元件。通过反相微乳液法制备近乎单分散的球形硅壳包覆的四氧化三铁（Fe3O4@SiO2）纳米粒子。通过硅壳与 3-溴丙基三氯硅烷的功能化，然后用叠氮化钠取代，制备叠氮功能化的 Fe3O4@SiO2。炔基化抗体和辣根过氧化物酶通过点击反应与叠氮功能化的 Fe3O4@SiO2 偶联作为信号标记。辣根过氧化物酶和四氧化三铁可以在过氧化氢存在下催化硫堇的氧化。在三明治免疫反应后，电流与微囊藻毒素-LR 的对数成正比。在 0.01 至 200 μg/mL 的范围内，线性回归方程为 i(μA)=119.89+46.27 log cMC-LR（μg/mL）。检测限为 0.004 μg/mL。该免疫分析方法将为环境监测和公共卫生提供一种通用的免疫分析方法。
