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

Dipak Kumar Rana1, Subhash Chandra Bhattacharya2

  • 1Department of Chemistry, Saldiha College, Bankura - 722173, West Bengal, India.

Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
|August 8, 2022
PubMed
Summary
This summary is machine-generated.

This study presents a novel pH-tunable Förster resonance energy transfer (FRET) sensor. The sensor utilizes Doxorubicin

Keywords:
BiosensingDoxorubicinExciplexFRET ON-OFFLifetimeLive cell imaging

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Area of Science:

  • Biochemistry and Molecular Biology
  • Materials Science
  • Nanotechnology

Background:

  • Förster resonance energy transfer (FRET) is a distance-dependent process crucial for studying molecular interactions.
  • Developing sensors with tunable emission properties, particularly pH-sensitive ones, is vital for biological and chemical applications.
  • Doxorubicin, an anticancer drug, exhibits pH-dependent optical properties that can be exploited for sensing.

Purpose of the Study:

  • To develop a novel, pH-tunable FRET system using Pyrazoline as a donor and Doxorubicin as an acceptor.
  • To investigate the pH-induced conformational changes in Doxorubicin and their effect on FRET efficiency.
  • To explore the potential of this FRET system as a sensor for intracellular pH and for controlled drug delivery.

Main Methods:

  • Spectroscopic analysis (UV-Vis absorption and fluorescence emission) to characterize FRET efficiency.
  • pH titration experiments to determine the optimal pH range for FRET.
  • Confocal fluorescence microscopy to visualize sensor performance in live HepG2 cells.

Main Results:

  • A tunable FRET emission was achieved by varying the medium's pH.
  • Maximum FRET efficiency was observed near the pKa of Doxorubicin due to optimal spectral overlap.
  • Abrupt ON-and-OFF FRET switching occurred due to pH-induced conformational changes in Doxorubicin, forming fluorescent exciplexes at elevated pH.

Conclusions:

  • The developed Pyrazoline-Doxorubicin system acts as an effective pH sensor with tunable FRET emission.
  • The sensor demonstrates efficient cell membrane penetration and potential for controlled drug delivery to tumor cells.
  • This approach offers a unique method for real-time monitoring of pH in biological systems.