Highly sensitive chemiluminescence biosensor for protein detection based on the functionalized magnetic microparticles and the hybridization chain reaction
Highly sensitive chemiluminescence biosensor for protein detection based on the functionalized magnetic microparticles and the hybridization chain reaction
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基于功能化磁性微粒和杂交链式反应的高灵敏蛋白质检测化学发光生物传感器
DOI：
10.1016/j.bios.2016.08.067
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发表时间：
2017-01-15
影响因子：
12.6
通讯作者：
He, Zhike
中科院分区：
文献类型：
--
作者：
Li, Ningxing;Chen, Jinyang;He, Zhike
基于功能化磁性微粒(MMPs)和杂交链式反应(HCR)，研制了一种用于蛋白质检测的超灵敏化学发光(CL)生物传感器。首先，将该引物与连接在MMPs表面的凝血酶适配子杂交。然后用部分底物引发HCR反应，并将产物组装到MMPs的表面。通过链霉亲和素与生物素的相互作用，将链霉亲和素-辣根过氧化物酶(SA-HRP)与HCR产物偶联。在凝血酶存在下，适体识别并结合到靶分子后，与SA-HRP结合的HCR产物从MMPs表面释放出来。因此，在加入H_2O_2-鲁米诺后，上清液中释放的SA-HRP产生了显著的化学发光成像信号。该方法对凝血酶的检出限可低至9.7fM。此外，通过改变试剂的加入顺序对传感策略进行了改进，并成功地应用于复杂样品中凝血酶的检测。此外，该方法还可以进行DNA检测，证明了该传感策略的普适性。(C)2016爱思唯尔B.V.保留所有权利。
An ultrasensitive chemiluminescence (CL) biosensor for the detection of protein is developed in this study based on the functionalized magnetic microparticles (MMPs) and the hybridization chain reaction (HCR). First, the primer hybridized with the thrombin aptamer conjugated on the surface of MMPs. Then the HCR was triggered by part of the primer and its products were assembled on the surface of the MMPs. Through the interaction between streptavidin and biotin, the streptavidin-horseradish peroxidase (SA-HRP) was coupled with the HCR products. In the presence of thrombin, the HCR products conjugating with SA-HRP were released from the surface of MMPs after the aptamer recognized and bound to its target molecule. So the released SA-HRP in the supernatant produced a significant chemiluminescence imaging signal after the addition of H2O2-luminol. The detection limit of thrombin with this method could be as low as 9.7 fM. Besides, the sensing strategy was modified by changing the adding order of reagents that was then successfully applied in the detection of thrombin in complex sample. What's more, the DNA detection also could be carried out with this method, which demonstrated the universality of the proposed sensing strategy. (C) 2016 Elsevier B.V. All rights reserved.