State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University , Changsha, Hunan 410082, China.
Anal Chem. 2014 Jul 1;86(13):6572-9. doi: 10.1021/ac501088q. Epub 2014 Jun 23.
An aptamer screening method using a positive and negative selection units integrated microfluidic chip was introduced. Here, myoglobin (Myo), one of the early markers to increase after acute myocardial infarction, was used as the model. After 7-round selection, the aptamers, which exhibited dissociation constants (K(d)) in the nanomolar range (from 4.93 to 6.38 nM), were successfully obtained using a positive and negative selection units integrated microfluidic chip. The aptamer with the highest affinity (K(d) = 4.93 nM) was then used for the fabrication of a label-free supersandwich electrochemical biosensor for Myo detection based on target-induced aptamer displacement. The detection limit of this aptamer-based electrochemical biosensor was 10 pM, which was significantly lower than that of those previous antibody-based biosensors for Myo detection. This work may not only develop a strategy for screening aptamer but also offer promising alternatives to the traditional analytical and immunological methods for Myo detection.
介绍了一种使用正、负筛选单元集成微流控芯片的适体筛选方法。在这里，肌红蛋白（Myo）被用作模型，它是急性心肌梗死后最早升高的标志物之一。经过 7 轮筛选，成功地从正、负筛选单元集成微流控芯片中获得了具有纳摩尔级解离常数（K(d)）的适体（4.93 至 6.38 nM）。然后，使用亲和力最高的适体（K(d) = 4.93 nM）构建了一种基于目标诱导适体置换的无标记超三明治电化学生物传感器，用于检测肌红蛋白。基于适体的电化学生物传感器的检测限为 10 pM，明显低于以前用于检测肌红蛋白的基于抗体的生物传感器。这项工作不仅可以开发适体筛选策略，还可以为传统的分析和免疫方法提供有前途的替代方案，用于肌红蛋白的检测。
