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Author
Sánchez Baygual, Francisco-Javier
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Abstract
Intensity-based fluorescent synthetic probes have been widely used to monitor and image a wide range of analytes in biological systems. Nevertheless, intensity-based probes have several drawbacks such as variations in probe environment or variations in probe concentration that may affect the reliability of the fluorescence measurement. Interestingly, ratiometric fluorescent probes can be used to overcome these problems.
In light of the above, in this final degree project, a Förster resonance energy transfer (FRET) based molecular probe for singlet oxygen (1O2) imaging and detection has been designed, synthesized and characterized. Proving that FRET-based platforms can be successfully used to detect singlet oxygen.
In a first step, a non-cleavable 1O2 probe (SN-011) and a cleavable 1O2 probe (SN-015) were synthesized by covalently attaching a fluorescein derivative (donor) and a Rhodamine B de-rivative (acceptor) to 3,6,9-trioxaundecanodioic acid yielding the non-cleavable control probe probe, SN-011 and to (Z)-3,3’-(ethene-1,2-diylbis(sulfanediyl))dipropanoic acid furnishing the cleavable probe (SN-015) which upon reaction with 1O2 linker cleavage provokes FRET dis-ruption.
In a second step, the photophysical properties and 1O2 detection performance of the synthe-sized probes were evaluated in solution trough fluorescence, absorption and time-resolved fluorescence measurements.
Lastly, 1O2 detection viability of SN-015 probe was evaluated in E. coli BL21 (DE3).
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