Key Laboratory of Marine Environmental Corrosion and Bio-fouling, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.
Open Studio for Marine Corrosion and Protection, Pilot National Laboratory for Marine Science and Technology (Qingdao), 1 Wenhai Road, Qingdao 266237, China.
Anal Methods. 2022 Jun 23;14(24):2350-2356. doi: 10.1039/d2ay00416j.
In this paper, we present a robust and versatile method for developing non-label aptasensors with high sensitivity. Amyloid-like protein aggregates were facilely synthesized with the commonly used passivating agent bovine serum albumin (BSA) in developing biosensors, and the produced amyloid-like phase-transited BSA (PTB) exhibited excellent antifouling performances and robust interfacial adhesion with the electrode surface. In order to improve the detection sensitivity of electrochemical measurements, reduced graphene oxide was electrochemically deposited onto the electrode surface. Moreover, gold nanoparticles were introduced to enhance the conductivity of the PTB film and facilitate subsequent aptamer modification. Two common biological species, adenosine triphosphate (ATP) and cytochrome c (cyt c), were chosen as detection targets, and their corresponding aptasensors were successfully constructed and systematically evaluated. The proposed aptasensors based on the PTB-Au antifouling composite exhibited high sensitivity and specificity towards ATP and cyt c detection, and the detection limits were calculated to be 0.26 nM and 0.64 nM for ATP and cyt c, respectively. Hence, this work provides a simple approach to develop highly sensitive aptasensors without any labeling process, and thus promises its great application in biological analysis.
在本文中，我们提出了一种稳健且通用的方法，用于开发具有高灵敏度的非标记适体传感器。在开发生物传感器时，我们使用常用的封闭剂牛血清白蛋白（BSA）方便地合成了类淀粉蛋白聚集物，所产生的类淀粉相转变的 BSA（PTB）表现出优异的抗污性能和与电极表面的强界面附着力。为了提高电化学测量的检测灵敏度，将还原氧化石墨烯电化学沉积到电极表面。此外，引入了金纳米粒子来提高 PTB 膜的导电性，并便于随后的适体修饰。选择两种常见的生物物种，三磷酸腺苷（ATP）和细胞色素 c（cyt c）作为检测目标，并成功构建和系统评估了相应的适体传感器。基于 PTB-Au 抗污复合材料的提出的适体传感器对 ATP 和 cyt c 的检测表现出高灵敏度和特异性，检测限分别计算为 0.26 nM 和 0.64 nM。因此，这项工作提供了一种简单的方法来开发无需任何标记过程的高灵敏度适体传感器，因此有望在生物分析中得到广泛应用。
