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Dalvit, C. et al.

Dalvit, Flocco, Veronesi, Stockman,

Fluorine-NMR competition binding experiments for high-throughput screening of large compound mixtures.

High-throughput ligand-based NMR screening with competition binding experiments is extended to (19)F detection. Fluorine is a favorable nucleus for these experiments because of the significant contribution of the Chemical Shift Anisotropy (CSA) to the (19)F transverse relaxation of the ligand signal when bound to a macromolecular target. A low to moderate affinity ligand containing a fluorine atom is used as a reference molecule for the detection and characterization of new ligands. Titration NMR experiments with the selected reference compound are performed for finding the optimal set-up conditions for HTS and for deriving the binding constants of the identified NMR hits. Rapid HTS of large chemical mixtures and plant or fungi extracts against the receptor of interest is possible due to the high sensitivity of the (19)F nucleus and the absence of overlap with the signals of the mixtures to be screened. Finally, a novel approach for HTS using a reference molecule in combination with a control molecule is presented.[1]

References

Fluorine-NMR competition binding experiments for high-throughput screening of large compound mixtures. Dalvit, C., Flocco, M., Veronesi, M., Stockman, B.J. Comb. Chem. High Throughput Screen. (2002) [Pubmed]

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