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Comparison of turn-on and ratiometric fluorescent G-quadruplex aptasensor approaches for the detection of ATP.
- Source :
-
Analytical and bioanalytical chemistry [Anal Bioanal Chem] 2019 Mar; Vol. 411 (7), pp. 1319-1330. Date of Electronic Publication: 2019 Jan 05. - Publication Year :
- 2019
-
Abstract
- Two fluorescent aptasensor methods were developed for the detection of ATP in biochemical systems. The first method consisted of a label-free fluorescent "turn-on" approach using a guanine-rich ATP aptamer sequence and the DNA-binding agent berberine complex. In the presence of ATP, the ATP preferentially binds with its aptamer and conformationally changes into a G-quadruplex structure. The association of berberine with the G-quadruplex results in the enhancement of the fluorescence signal of the former. The detection limit of ATP was found to be 3.5 μM. Fluorescence, circular dichroism and melting temperature (T <subscript>m</subscript> ) experiments were carried out to confirm the binding specificity and structural changes. The second method employs the ratiometric fluorescent approach based on the Forster resonance energy transfer (FRET) for the detection of ATP using berberine along with a quencher (AuNRs, AgNPs) and a fluorophore (red quantum dots (RQDs), carbon dots (CDs)) labeled at 5' and 3' termini of the ATP-binding aptamer sequence. Upon addition of ATP and berberine, ATP specifically binds with its aptamer leading to the formation of G-quadruplex, and similarly, berberine also binds to the G-quadruplex. This leads to an enhancement of fluorescence of berberine while that of RQD and CDs were significantly quenched via FRET. The respective detection limits calculated were 3.6 μM and 3.8 μM, indicating these fluorescent aptasensor methods may be used for a wide variety of small molecules. Graphical abstract.
- Subjects :
- Adenosine Triphosphate analysis
Berberine chemistry
Fluorescence
Fluorescence Resonance Energy Transfer methods
Guanine chemistry
Humans
Limit of Detection
Quantum Dots chemistry
Adenosine Triphosphate blood
Aptamers, Nucleotide chemistry
Biosensing Techniques methods
Fluorescent Dyes chemistry
G-Quadruplexes
Subjects
Details
- Language :
- English
- ISSN :
- 1618-2650
- Volume :
- 411
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Analytical and bioanalytical chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 30612178
- Full Text :
- https://doi.org/10.1007/s00216-018-1484-x