Differentiation of Folate-Receptor-Positive and -Negative Cells Using a Substrate-Mimicking Fluorescent Probe

Diagnosis and therapy exploiting overexpressed receptors on the cell surface is one important strategy in medicine. Determination of the over expression level of a particular receptor is prerequisite for it to be of clinical use. Differentiation between FR-positive (FR=folate receptor) and -negative cells via fluorescence microscopy using a substrate mimetic fluorophore is presented in this work. The strategy adopted here is not the classical FA-conjugated (FA=folic acid) fluorescent probe but a small and environment-sensitive pterin-based (pterin is part of folate, i.e., vitamin B9) fluorescent probe. Electronically diverse pterin-based fluorescent probes have been designed and synthesized to understand the effect of the binding environment on the receptor-substrate interactions. By utilizing steady-state UV/Vis and fluorescence along with time-resolved fluorescence spectroscopy, the effects on the electronic and acid-base properties of the substrate were investigated. Evidently, one synthesized probe showed FA-mimicking behavior with strong binding interaction with FR.