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Selection of a DNA aptamer that binds 8-OHdG using GMP-agarose
Authors:Yusuke Miyachi  Nobuaki Shimizu  Chiaki Ogino  Hideki Fukuda  Akihiko Kondo
Affiliation:1. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Rokkoudai-chou 1-1, Nada, Kobe 657-8501, Japan;2. Division of Material Sciences, Graduate School of Neutral Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan
Abstract:DNA aptamers, which bind specific molecule, such as 8-OHdG, with high affinity were investigated using an in vitro selection strategy called systematic evolution of ligands by exponential enrichment (SELEX). However, 8-OHdG was difficult to immobilize on a carrier for SELEX. Therefore, a DNA aptamer binding to 8-OHdG was selected using GMP-agarose as an analogue from a library of about 460 random ssDNA sources. As a result, three aptamer candidates were selected. Among the selected DNA aptamers, the No. 22 DNA aptamer exhibited a high affinity for 8-OHdG. The dissociation constant, KD, of No. 22 DNA aptamer was on the order of 0.1 μmol/L. This result suggests that using an analogue will be a useful new SELEX method for obtaining various aptamers that are difficult to immobilize on a matrix.
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