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Analysis of mismatched DNA by mismatch binding ligand (MBL)–Sepharose affinity chromatography

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Abstract

Mismatch binding molecules (MBLs), strongly and selectively bound to the mismatched base pair in duplex DNA, were immobilized on Sepharose. Three MBL–Sepharose columns were prepared with three MBLs, naphthyridine dimer (ND), naphthyridine–azaquinolone (NA), and aminonaphthyridine dimer (amND), which exhibited different binding profiles to the mismatched base pairs. These three MBL–Sepharose columns showed characteristic elution profiles for DNA duplexes containing mismatched base pairs. The ND–Sepharose column separated the G–G and G–A mismatched DNA from fully matched duplexes. The NA–Sepharose column separated the A–A and G–A mismatched DNA from other DNA duplexes. The amND–Sepharose column separated the C–C mismatched DNA. These chromatographic profiles were very consistent with the binding preference of each MBL. By changing the elution conditions from sodium hydroxide to sodium chloride, MBL–Sepharose columns were also able to separate the mismatched DNA that weakly bound to the MBL from fully matched DNA duplex.

MBL-Sepharose affinity chromatography successfully separates the mismatched duplex DNA from fully matched duplex.

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Author notes

  1. Kazuhiko Nakatani

    Present address: The Institute of Scientific and Industrial Research (SANKEN), Osaka University, 8-1, Mihogaoka, Ibaraki, 567-0047, Japan

Authors and Affiliations

  1. Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, 615-8510, Japan

    Yuki Goto, Hitoshi Suda, Akio Kobori & Kazuhiko Nakatani

Authors
  1. Yuki Goto
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  2. Hitoshi Suda
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  3. Akio Kobori
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  4. Kazuhiko Nakatani
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Correspondence to Kazuhiko Nakatani.

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Goto, Y., Suda, H., Kobori, A. et al. Analysis of mismatched DNA by mismatch binding ligand (MBL)–Sepharose affinity chromatography. Anal Bioanal Chem 388, 1165–1173 (2007). https://doi.org/10.1007/s00216-007-1323-y

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  • DOI: https://doi.org/10.1007/s00216-007-1323-y

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