Development of RNA aptamer and its ligand binding assay on microchip electrophoresis.
Development of RNA aptamer and its ligand binding assay on microchip electrophoresis.
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DOI：
10.2174/1875397301206010001
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Arakawa H
中科院分区：
文献类型：
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作者：
Ohno K;Nakata C;Sano Y;Nishikawa F;Nishikawa S;Arakawa H
采用微芯片电泳结合荧光检测技术，通过指数富集（SELEX）轮对靶分子进行配体系统进化，评价适体的结合活性。这种方法是一种非放射性同位素、快速和简单的平台，并且电泳分离似乎是用于寡核苷酸分子的适体的有效技术。我们尝试通过上述方法获得促性腺激素特异性RNA适体。结果，基于它们之间的构象差异的适体的峰被分离并在电泳图上检测到。此外，未结合适体的峰强度随着通过SELEX轮的进展而降低，表明通过所提出的方法获得具有高亲和力的RNA适体。
Microchip electrophoresis (ME) coupled with fluorescence detection was used to estimate the binding activity of aptamer in each systematic evolution of ligands by exponential enrichment (SELEX) round for a target molecule. This approach is a non-radioisotopic, rapid and simple platform, and electrophoretic separation appears to be an effective technique for aptamers of oligonucleotide molecules. We tried to obtain gonadotropin-specific RNA aptamer by the above approach. As a result, the peaks of aptamers based on the conformational differences between them were separated and detected on the electropherograms. Moreover, the intensity of peak of unbound aptamer was decreased with progression through the SELEX rounds, suggesting that RNA aptamer with high affinity was obtained by the proposed method.