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

Goto, Yuki; Suda, Hitoshi; Kobori, Akio; Nakatani, Kazuhiko
July 2007
Analytical & Bioanalytical Chemistry;Jul2007, Vol. 388 Issue 5/6, p1165
Academic Journal
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. [Figure not available: see fulltext.]


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