RESEARCH ARTICLE


Development of RNA Aptamer and its Ligand Binding Assay on Microchip Electrophoresis



Ken-ichi Ohnoa, Chikara Nakataa, Yoshihiro Sanoa, Fumiko Nishikawab, Satoshi Nishikawab, Hidetoshi Arakawa*, a
a School of Pharmacy Showa University, 1-5-8 Hatanodai, Shinagawa, Tokyo 142-8555, Japan
b Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan


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© Ohno et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the School of Pharmacy, Showa University, 1-5-8 Hatanodai, Shinagawa, Tokyo 142-8555, Japan; Tel: +81-3-3784-8194; Fax: +81-3-3784-8247; E-mail: arakawa@pharm.showa-u.ac.jp


Abstract

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.

Keywords: Aptamer, conformational differences, fluorescence detection, intercalating dyes, microchip electrophoresis, SELEX.