Apoferritin protein nanoparticles dually labeled with aptamer and horseradish peroxidase as a sensing probe for thrombin detection.
Apoferritin protein nanoparticles dually labeled with aptamer and horseradish peroxidase as a sensing probe for thrombin detection.
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DOI：
10.1016/j.aca.2012.10.041
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发表时间：
2013-01
影响因子：
6.2
通讯作者：
Jie Zhao;Meiling Liu;Youyu Zhang;Haitao Li;Yuehe Lin;S. Yao
中科院分区：
文献类型：
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作者：
Jie Zhao;Meiling Liu;Youyu Zhang;Haitao Li;Yuehe Lin;S. Yao
基于HRP和脱铁铁蛋白的双重信号放大作用，开发了一种新型的超灵敏的HRP型电化学适体传感器用于凝血酶的检测。以核/壳Fe 3 O 4/Au磁性纳米粒子（AuMNP）负载适配子1（Apt 1）作为识别元件，以Apt 2和HRP双标记的脱铁铁蛋白（Apt 2 + HRP）作为检测探针。Apt 1/凝血酶/Apt 2-脱铁铁蛋白NPs-HRP是通过AuMNPs-Apt 1、凝血酶和Apt 2-脱铁铁蛋白-HRP之间的亲和反应形成的三明治型复合物。将复合物锚定在丝网印刷碳电极（SPCE）上。采用微分脉冲伏安法（DPV）监测电极响应。所提出的适体传感器在0.5- 100 pM的宽范围内对凝血酶浓度产生线性电流响应，检测限为0.07pM（S/N=3）。用脱铁铁蛋白和HRP对检测信号进行放大。这种基于纳米颗粒的适体传感器提供了一种快速、灵敏、选择性和廉价的凝血酶定量的新方法，并在蛋白质检测和疾病诊断中提供了有前途的潜力。
A novel and ultrasensitive sandwich-type electrochemical aptasensor has been developed for the detection of thrombin, based on dual signal-amplification using HRP and apoferritin. Core/shell Fe3O4/Au magnetic nanoparticles (AuMNPs) loading aptamer1 (Apt1) was used as recognition elements, and apoferritin dually labeled with Aptamer2 (Apt2) and HRP was used as a detection probe. Sandwich-type complex, Apt1/thrombin/Apt2–apoferritin NPs–HRP was formed by the affinity reactions between AuMNPs–Apt1, thrombin, and Apt2–apoferritin–HRP. The complex was anchored on a screen-printed carbon electrode (SPCE). Differential pulse voltammetry (DPV) was used to monitor the electrode response. The proposed aptasensor yielded a linear current response to thrombin concentrations over a broad range of 0.5–100pM with a detection limit of 0.07pM (S/N=3). The detection signal was amplified by using apoferritin and HRP. This nanoparticle-based aptasensor offers a new method for rapid, sensitive, selective, and inexpensive quantification of thrombin, and offers a promising potential in protein detection and disease diagnosis.