Novel Förster Resonance Energy Transfer probe with quantum dot for a long-time imaging of active caspases inside individual cells

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Authors

PROCHÁZKOVÁ Markéta KUCHOVSKÁ Eliška KILLINGER Michael KLEPÁRNÍK Karel

Year of publication 2023
Type Article in Periodical
Magazine / Source Analytica Chimica Acta
MU Faculty or unit

Faculty of Science

Citation
Web https://www.sciencedirect.com/science/article/pii/S000326702300555X
Doi http://dx.doi.org/10.1016/j.aca.2023.341334
Keywords Caspase molecular probe; Time-laps fluorescence microscopy; Quantum dot photostability; FRET probe
Description With the goal to investigate biological phenomena at a single-cell level, we designed, synthesized and tested a molecular probe based on Förster resonance energy transfer (FRET) between a highly luminescent quantum dot (QD) as a donor and a fluorophore or fluorescence quencher as an acceptor linked by a specific peptide. In principle, QD luminescence, effectively dissipated in the probe, is switched on after the cleavage of the peptide by a protease and the release of the quencher. We proposed a novel synthesis strategy of a probe. A two-step synthesis consists of: (i) Conjugation of CdTe QDs functionalized by –COOH groups of succinic acid on the nanoparticle surface with the designed specific peptide (GTADVEDTSC) using a ligand-exchange approach; (ii) A fast, high-yield reaction of amine-reactive succinimidyl group on the BHQ-2 quencher with N-terminal of the peptide.
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