Modified capillary electrophoresis based measurement of the binding between DNA aptamers and an unknown concentration target

文献类型： 外文期刊

第一作者： Zhang, Yue-Wei

作者： Zhang, Yue-Wei;Yan, Hai-Yu;Fu, Ping;Jiang, Fei;Zhang, Yao;Wu, Wen-Xue;Li, Jin-Xiang

作者机构：

关键词： Aptamer;Capillary electrophoresis;Complicated target;Dissociation constant

期刊名称：ANALYTICAL AND BIOANALYTICAL CHEMISTRY （ 影响因子：4.142； 五年影响因子：3.863 ）

ISSN：

年卷期：

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收录情况： SCI

摘要： The recognition of targets such as biomacromolecules, viruses and cells by their aptamers is crucial in aptamer-based biosensor platforms and research into protein function. However, it is difficult to evaluate the binding constant of aptamers and their targets that are hard to purify and quantify, especially when the targets are undefined. Therefore, we aimed to develop a modified capillary electrophoresis based method to determine the dissociation constant of aptamers whose targets are hard to quantify. A protein target, human thrombin, and one of its aptamers were used to validate our modified method. We demonstrated that the result calculated by our method, only depending on the aptamer's concentrations, was consistent with the classical method, which depended on the concentrations of both the aptamers and the targets. Furthermore, a series of DNA aptamers binding with avian influenza virus H9N2 were confirmed by a four-round selection of capillary electrophoresis-systematic evolution of ligands by exponential enrichment, and we identified the binding constant of these aptamers by directly using the whole virus as the target with the modified method. In conclusion, our modified method was validated to study the interaction between the aptamer and its target, and it may also advance the evaluation of other receptor-ligand interactions.

分类号： O65

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