Chimeric Aptamers-Based and MoS₂ Nanosheet-Enhanced Label-Free Fluorescence Polarization Strategy for Adenosine Triphosphate Detection

Abstract

Adenosine triphosphate (ATP) as a primary energy source plays a unique role in the regulation of all cellular events. The necessity to detect ATP requires sensitive and accurate quantitative analytical strategies. Herein, we present our study of developing a MoS 2 nanosheet-enhanced aptasensor for fluorescence polarization-based ATP detection. A bifunctional DNA strand was designed to consist of chimeric aptamers that recognize and capture ATP and berberine, a fluorescence enhancer. In the absence of ATP, the DNA strand bound to berberine will be hydrolyzed when Exonuclease I (Exo I) is introduced, releasing berberine as a result. In contrast, when ATP is present, ATP aptamer folds into a G-quadruplex structure; thus, the complex can resist degradation by Exo I to maintain berberine for fluorescent detection purpose. In addition, to magnify the fluorescence polarization (FP) signal, MoS 2 nanosheets were also adopted in the system. This nanosheets-enhanced FP strategy is simple and facile which does not require traditional dye-labeled DNA strands and complex operation steps. The developed fluorescence polarization aptasensor showed high sensitivity for the quantification of ATP with a detection limit of 34.4 nM, performing well both in buffer solution and in biological samples.

Ex vivo and in vivo fluorescence detection and imaging of adenosine triphosphate

“…The serum and urine samples were extracted from mouse or human. ATP solutions covering from 0.0 to 1.0 mM were separately spiked into the 0.1% diluted three different body fluid samples (e.g., mouse serum, mouse urine, and human serum), respectively [ 10 ]. Finally, the fluorescence intensity at 610 nm of the TC/Apt-based fluorescent probes could be detected and calculated from a series of PL spectra of ROX dyes after adding 200 μg/mL TC/Apt probes into above three different body fluid samples containing the ATP with various concentrations.…”

Ex Vivo and In Vivo Fluorescence Detection and Imaging of Adenosine Triphosphate

“…The serum and urine samples were extracted from mouse or human. ATP solutions covering from 0.001 to 5.0 mM were separately spiked into the 0.1% diluted three different body uid samples (e.g., mouse serum, rat urine, and human serum), respectively 10 . Finally, the uorescence intensity at 610 nm of the TC/Apt-based uorescent probes could be detected and calculated from a series of PL spectra of ROX dyes after adding 200 µg/mL TC/Apt probes into above three different body uid samples containing the ATP with various concentrations.…”

“…To this end, numerous approaches have been developed for ATP detection, including surface-enhanced Raman scattering, electrochemistry, chemiluminescence, colorimetry, uorescence and so on [8][9][10][11][12][13][14][15][16] . Among these methods, uorescent probes for imaging and/or sensing intracellular ATP variations have attracted enormous attention due to their high sensitivity, good selectivity, convenient measurement and low cost 1,[17][18][19][20] .…”

Two-dimensional nanomaterials for Förster resonance energy transfer–based sensing applications

“…The LOD of the MoS 2 -based assay was calculated to be 500 pm and 5 μm for DNA and adenosine, respectively. Since then, dozens of researchers have reported the development of MoS 2 -DNA-based FRET sensors for the detection of various targets, such as the nucleic acids [74][75][76][77][78][79][80][81][82][83][84][85], proteins [86][87][88][89][90][91][92][93][94][95], metal ions [96,97], small molecules [52,86,[98][99][100][101][102][103][104], enzyme activity [105], and others [106]. These sensors mainly take advantage of the fluorescence-quenching capability and different affinity of MoS 2 to ssDNA and ssDNA-target combination, enabling easy preparation, single-step, and rapid detection.…”