Quantum dots-based fluorescence resonance energy transfer biosensor for monitoring cell apoptosis
Abstract
The development of advanced methods for accurately monitoring cell apoptosis has extensive significance in the diagnostic and pharmaceutical fields. In this study, we developed a rapid, sensitive and selective approach for the detection of cell apoptosis by combining the site-specific recognition and cleavage of the DEVD-peptide with quantum dots (QDs)-based fluorescence resonance energy transfer (FRET). Firstly, biotin-peptide was conjugated on the surface of AuNPs to form AuNPs-pep through the formation of an Au-S bond. Then, AuNPs-pep-QDs nanoprobe was obtained through the connection between AuNPs-pep and QDs. FRET is on and the fluorescence of QDs is quenched at this point. The evidence of UV-vis spectra, transmission electron microscopy (TEM), and Fourier transform infrared (FT-IR) spectroscopy revealed that the connection was successful. Upon the addition of apoptosis cell lysis solution, peptide was cleaved by caspase-3, and AuNPs was dissociated from the QDs. At this time, FRET is off, and thus the QDs fluorescence was recovered. The experimental conditions were optimized in terms of ratio of peptide to AuNPs, buffer solution, and the temperature of conjugation and enzyme reaction. The biosensor was successfully applied to distinguishing apoptosis cells and normal cells within 2 h. This study demonstrated that the biosensor could be utilized to evaluate anticancer drugs.
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