Analytical Sciences

Identification of Binding Epitopes of a Fluorinated Compound Bound to Proteins Using ¹H and ¹⁹F NMR Spectroscopy

Kazuo FURIHATA* and Mitsuru TASHIRO**

*Division of Agriculture and Agricultural Life Sciences, The University of Tokyo, Yayoi, Bunkyo, Tokyo 113-8657, Japan
**Department of Chemistry, College of Science and Technology, Meisei University, Hino, Tokyo 191-8506, Japan

¹H/¹⁹F NMR-based screening methods were applied to a human serum albumin-fleroxacin complex. Fleroxacin contains three fluorine atoms in a molecule, which is suitable as a model fluorinated compound for NMR analysis with ¹H and ¹⁹F detection. The ¹⁹F{¹H} and ¹H{¹H} saturation transfer difference spectra were acquired and the ¹H/¹⁹F spin-lattice relaxation rates were measured with and without any selective irradiation of protein resonance to identify the binding epitopes of fleroxacin. Because several ¹H signals of fleroxacin resonated close to water, its precise signal intensities were unavailable. The ¹⁹F NMR-based screening methods successfully provide complementary information, indicating its importance in the analysis of fluorinated compounds.

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