Department of Reproductive Genetics, Women's Hospital Zhejiang University School of Medicine, Hangzhou, 310006, Zhejiang Province, China.
Department of Central Laboratory, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, 310006, Zhejiang Province, China.
Mikrochim Acta. 2019 Mar 30;186(4):262. doi: 10.1007/s00604-019-3359-z.
An ultrasensitive sandwich-type electrochemical immunosensor was developed for the amperometric determination of serum myeloperoxidase (MPO). The method is making use of (a) gold nanoparticles encapsulated in graphitized mesoporous carbons (AuNP@GMC); and (b) horseradish peroxidase (HRP) labeled secondary antibody (HRP@Ab) immobilized on AuNP@GMC. MPO capture antibody (Ab) was immobilized on the electrode modified with an AuNP-graphene oxide nanocomposite. The sandwich immunoreaction leads to the formation of the complex composed of Ab, MPO, and HRP@Ab. An amplified electrochemical signal is produced by electrocatalytic reduction of HO (at a typical voltage of -0.18 V vs. Ag/AgCl) in the presence of enzymatically oxidized thionine. The peak current of thionine was measured using differential pulse voltammetry. Under optimized steady-state conditions, the reduction peak increases in the 1 to 300 pg.mL MPO concentration range, and the detection limit is 0.1 pg.mL (at S/N = 3). Graphical abstract Schematic presentation of AuNP-GO based sandwich-type electrochemical immunoassay for the determination of myeloperoxidase by using gold nanoparticles encapsulated in graphitized mesoporous carbons (AuNP@GMC) as a carrier for horseradish peroxidase (HRP) labeled secondary antibody (HRP@Ab).
一种超灵敏的三明治型电化学免疫传感器被开发用于血清髓过氧化物酶（MPO）的电流测定。该方法利用（a）封装在石墨化介孔碳中的金纳米粒子（AuNP@GMC）；和（b）固定在 AuNP@GMC 上的辣根过氧化物酶（HRP）标记的二级抗体（HRP@Ab）。MPO 捕获抗体（Ab）被固定在修饰有 AuNP-氧化石墨烯纳米复合材料的电极上。三明治免疫反应导致由 Ab、MPO 和 HRP@Ab 组成的复合物的形成。在存在酶促氧化噻嗪的情况下，通过 HO 的电化学催化还原（相对于 Ag/AgCl 的典型电压为-0.18 V）产生放大的电化学信号。使用差分脉冲伏安法测量噻嗪的峰电流。在优化的稳态条件下，还原峰在 1 到 300 pg.mL MPO 浓度范围内增加，检测限为 0.1 pg.mL（在 S/N = 3 时）。
