Control of electrostatic interaction between a molecular beacon aptamer and conjugated polyelectrolyte for detection range-tunable ATP assayElectronic supplementary information (ESI) available: Experimental details, physical and optical properties of CPEs and additional spectra and figures. See DOI: 10.1039/c7py01252g

A new strategy to modulate the detection range of a molecular beacon aptamer (MBA)/conjugated polyelectrolyte (CPE)-based adenosine triphosphate (ATP) assay is suggested. The hairpin type probe, MBA contains the ATP-specific aptamer sequence in a loop part and a green fluorophore (6-carboxyfluoresce...

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Hauptverfasser: Jeong, J.-E, Woo, H. Y
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Sprache:eng
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Zusammenfassung:A new strategy to modulate the detection range of a molecular beacon aptamer (MBA)/conjugated polyelectrolyte (CPE)-based adenosine triphosphate (ATP) assay is suggested. The hairpin type probe, MBA contains the ATP-specific aptamer sequence in a loop part and a green fluorophore (6-carboxyfluorescein, 6-FAM) and quencher (4-((4-(dimethylamino)phenyl)azo)benzoic acid) at both termini of stem parts. Three kinds of water-soluble poly(fluorene-phenylene) based CPEs ( MP2-MP6 ) were synthesized by varying the number of cationic groups (2 to 6) per repeat unit. In the absence of ATP, the hairpin MBA is transformed into open-chain conformation by forming a linear electrostatic complex with CPE, enabling facile fluorescence resonance energy transfer from blue-emitting CPEs to 6-FAM in MBA. In the presence of ATP, MBA forms a G-quadruplex with ATP where the close contact between the fluorophore and quencher results in the quenched PL signal of 6-FAM. The binding interaction between the aptamer and ATP in the G-quadruplex shows resistance to the structural opening by CPEs and the ionic density of CPEs was proved to have an important role in the structural transformation of the ATP-MBA G-quadruplex into the linear open-chain MBA/CPE complex. By increasing the ionic density of CPEs, structural transformation toward open-chain MBA/CPE complexes was accelerated due to enhanced electrostatic interaction between the MBA and CPE, resulting in a detection range shift to a higher [ATP]. By simply modifying the number of ionic groups in CPE, the population shift toward open-chain MBA/CPE demonstrates the successful tuning of detection range from nanomolar up to millimolar concentrations of ATP. A new strategy is suggested to fine-tune the detection range by controlling the ionic density of CPEs in the MBA/CPE-based ATP assay.
ISSN:1759-9954
1759-9962
DOI:10.1039/c7py01252g