Implication toward a simple strategy to generate pH tunable FRET-based biosensing

The present contribution depicts a unique approach to generate tunable Förster resonance energy transfer (FRET) emission with variation of pH of the medium. The pH sensitive absorption of Doxorubicin leads to modification of spectral overlap between emission spectra of donor (Pyrazoline) and absorption spectra of acceptor (Doxorubicin) thereby sensing maximum FRET efficiency in an optimum pH (near pK_a of Doxorubicin). This drug molecule exhibits an instantaneous conformation change at a particular pH, which consequences on abrupt ON-and-OFF FRET efficiency. At elevated pH, both the drug molecules exhibit conformational change and form stable fluorescent exciplex, switching off the FRET emission. Confocal fluorescence images of live HepG2 cells imply that the sensor can proficiently go through the cell membrane and can be applied in the controlled delivery of drug to the tumor cell lines.