Highly sensitive electrochemical nuclear factor kappa B aptasensor based on target-induced dual-signal ratiometric and polymerase-assisted protein recycling amplification strategy.
Highly sensitive electrochemical nuclear factor kappa B aptasensor based on target-induced dual-signal ratiometric and polymerase-assisted protein recycling amplification strategy.
复制标题
DOI：
10.1016/j.bios.2017.10.048
复制
发表时间：
2018-04
影响因子：
12.6
通讯作者：
Kanfu Peng;Pan Xie;Zhe-han Yang;R. Yuan;Keqin Zhang
中科院分区：
文献类型：
--
作者：
Kanfu Peng;Pan Xie;Zhe-han Yang;R. Yuan;Keqin Zhang
本工作基于靶结合触发的比率信号读出和聚合酶辅助的蛋白循环扩增策略，建立了一种用于核因子κ B（NF-κB）检测的放大电化学比率适体传感器。为了证明双信号电化学比率读数的“信号关闭”和“信号开启”变化的影响，巯基发夹DNA（SH-HD）与亚甲基蓝（MB）修饰的保护DNA（MB-PD）杂交以形成捕获探针，其被合理地引入用于构建测定平台。在界面上，探针可以特异性地结合靶NF-κB并暴露一个立足点区域，该立足点区域随后与二茂铁（Fc）修饰的DNA链杂交以将Fc基团带到电极表面，同时置换MB-PD以从电极表面释放MB基团，导致Fc（IFc）的“信号开启”和MB（IMB）的“信号关闭”。为了提高电化学适体传感器的灵敏度，设计了phi 29辅助的靶蛋白再循环扩增策略，以实现放大的比率信号。由于上述优点，所制备的适体传感器表现出0.1 pg mL− 1至15 ng mL− 1的宽线性范围，检测限为0.03 pg mL−1。该策略提供了一种简单而巧妙的方法来构建比率电化学适体传感器，并通过改变识别探针在多种疾病标志物检测中显示出有前途的潜在应用。
In this work, an amplified electrochemical ratiometric aptasensor for nuclear factor kappa B (NF-κB) assay based on target binding-triggered ratiometric signal readout and polymerase-assisted protein recycling amplification strategy is described. To demonstrate the effect of “signal-off” and “signal-on” change for the dual-signal electrochemical ratiometric readout, the thiol-hairpin DNA (SH-HD) hybridizes with methylene blue (MB)-modified protection DNA (MB-PD) to form capture probes, which is rationally introduced for the construction of the assay platform. On the interface, the probes can specifically bind to target NF-κB and expose a toehold region which subsequently hybridizes with the ferrocene (Fc)-modified DNA strand to take the Fc group to the electrode surface, accompanied by displacing MB-PD to release the MB group from the electrode surface, leading to the both “signal-on” of Fc (IFc) and “signal-off” of MB (IMB). In order to improve the sensitivity of the electrochemical aptasensor, phi29-assisted target protein recycling amplification strategy was designed to achieve an amplified ratiometric signal. With the above advantages, the prepared aptasensor exhibits a wide linear range of 0.1 pg mL−1to 15 ng mL−1with a low detection limit of 0.03 pg mL−1. This strategy provides a simple and ingenious approach to construct ratiometric electrochemical aptasensor and shows promising potential applications in multiple disease marker detection by changing the recognition probe.