Biochemists are investigating detection methods for living substances such as protein, and neurotransmitters. In this study, we have designed and synthesized three fluorescent molecular probes and performed the highly sensitive and selective detections of living substances using magnetic beads; i) fluorescent reagent using FRET between dansyl fluorophore as energy donor and cyanopyranyl group as energy acceptor for the detection of glycoconjugates (Probe 1) in combination with magnetic beads, ii) fluorescent molecule for the highly sensitive and selective detection of neurotransmitter (Probe 2), iii) fluorescent molecule for the highly sensitive and selective detection of vascular endothelial growth factor (VEGF) (Probe 3). In order to detect lectin-saccharide interactions using a novel FRET pair (Probe 1), the dansyl fluorophore and lectins were immobilized on the surface of magnetic particles, and the detections of various saccharide containing cyanopyranyl moiety were successfully performed in highly sensitive way. Moreover, this analytical system performed a highly-sensitive detection of glycoconjugates with high selectivity. A novel oxytocin-targeting fluorescent peptide (probe 2), with a cyanopyranyl group as the fluorophore and a peptide as the binding site of oxytocin, was developed. Probe 2 allowed the detection of oxytocin by alteration of its fluorescence quantum yield. When alone in solution and unbound to oxytocin, probe 2 on the surface of magnetic beads had a weak fluorescence emission, contrary to a much stronger fluorescence emission when associated with oxytocin. To take advantage of the molecular design of probe 2, probe 3 has been developed for the detection of VEGF, whose detection limit was 2.0 pg/mL. These results proved that above two fluorescent probes could be useful for a convenient and highly sensitive detections of biological substances in biochemical and medical fields.

Development of fluorescent molecular probes for the highly sensitive and selective detection of living substances using magnetic beads

2019 Fall National Meeting

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