Molecular Science and Biomedicine Laboratory, State Key Laboratory for Chemo/Bio Sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, and Collaborative Research Center of Molecular Engineering for Theranostics, Hunan University, Changsha 410082, China.
Attribute Sciences, Amgen, One Amgen Center Drive, Thousand Oaks, CA 91320, United States.
Talanta. 2017 Mar 1;164:662-667. doi: 10.1016/j.talanta.2016.11.001. Epub 2016 Nov 3.
In this paper, based on reciprocal chiral substrate specificity, taking achiral molecules, ethanolamine (EA) and malachite green (MG) as two model targets, biostable L- DNA aptamers and L-RNA aptamers were generated respectively by chiral inversion of existing D-aptamers. In the detection of EA with L-DNA aptamer-based sensors, the feasibility of our strategy was confirmed, while in the detection of MG with L-RNA aptamers, linear calibration curves were obtained in the range from 0.1 to 5µm with the detection limit of 0.065µm under optimized experimental conditions. The results demonstrated that the mirror-image L-aptamers have identical recognition capability as D-aptamers. Meanwhile, L-aptamers have superior biostability to resist nuclease digestion, protein binding interference and off-target effects, enabling their applications in complex practical samples, such as lake water and fish tissue extractions. Our work provides a simple, yet universal and efficient way to develop biostable aptamers.
在本文中，基于互反手性底物特异性，以非手性分子乙醇胺（EA）和孔雀石绿（MG）为两个模型靶标，通过对手性 D-适体进行手性反转，分别生成了稳定的 L-DNA 适体和 L-RNA 适体。在基于 L-DNA 适体的传感器检测 EA 的实验中，验证了我们策略的可行性，而在 L-RNA 适体检测 MG 的实验中，在优化的实验条件下，获得了 0.1 到 5µm 范围内的线性校准曲线，检测限为 0.065µm。结果表明，镜像 L-适体与 D-适体具有相同的识别能力。同时，L-适体具有优越的生物稳定性，能够抵抗核酸酶消化、蛋白质结合干扰和脱靶效应，使其能够应用于复杂的实际样品，如湖水和鱼组织提取。我们的工作为开发稳定的适体提供了一种简单、通用且高效的方法。
