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In vitro selection of sialic acid specific RNA aptamer and its application to the rapid sensing of sialic acid modified sugars

Authors

  • Suhyung Cho,

    1. Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, Republic of Korea; telephone: +82-2-880-6774; fax: +82-2-883-6020
    2. Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea
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  • Bo-Rahm Lee,

    1. School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, Korea
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  • Byung-Kwan Cho,

    1. Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea
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  • June-Hyung Kim,

    1. Department of Chemical Engineering, Dong-A University, Busan, Republic of Korea
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  • Byung-Gee Kim

    Corresponding author
    1. Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, Republic of Korea; telephone: +82-2-880-6774; fax: +82-2-883-6020
    2. School of Chemical and Biological Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, Korea
    • Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, Republic of Korea; telephone: +82-2-880-6774; fax: +82-2-883-6020
    Search for more papers by this author

Abstract

Sialic acids (SAs) are located on the terminal positions of glycan on a cell surface, which play important role in the spread and metastasis of cancer cells and infection of pathogen. For their detection and diagnosis, the finding of SA specific ligand is an essential prerequisite. Here, RNA aptamer for N-acetylneuraminic acid (Neu5Ac), a representative of SAs, with the high affinity of 1.35 nM and the selectivity was screened by in vitro selection method. The strong binding of the screened aptamer was enough to protect the hydrolysis of Neu5Ac by neuraminidase with the stoichiometry of 1:1 molar ratio. For the rapid detection of SAs, the RNA aptamer was further engineered to the aptazyme sensor by conjugating with a ribozyme following the characterization of selected aptamer by RNase footprinting assay. Without additional desialylation, modification, or/and purification processes, the aptazyme indicated high catalytic activities in the presence of Neu5Ac over 20 µM in several minutes. Also, we observed that the aptazyme sensor shows high sensitivities to Neu5Ac-conjugated sugars as well as Neu5Ac monomer, but not in non-Neu5Ac modified sugars. The aptamer for Neu5Ac can support valuable tools in a wide range of bioanalytical applications as well as biosensors. Biotechnol. Bioeng. 2013; 110: 905–913. © 2012 Wiley Periodicals, Inc.

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