Detection Of Adenosine Triphosphate Research Articles

Proximity-enabled bidirectional enzymatic repairing amplification for ultrasensitive fluorescence sensing of adenosine triphosphate

A novel fluorescence sensing strategy for ultrasensitive and highly specific detection of adenosine triphosphate (ATP) has been developed by the combination of the proximity ligation assay with bidirectional enzymatic repairing amplification (BERA). The strategy relies on proximity binding-triggered the release of palindromic tail that initiates bidirectional cyclic enzymatic repairing amplification reaction with the aid of polymerase and two DNA repairing enzymes, uracil-DNA glycosylase (UDG) and endonuclease IV (Endo IV). A fluorescence-quenched hairpin probe with a palindromic tail at the 3′ end is skillfully designed that functions as not only the recognition element, primer, and polymerization template for BERA but also the indicator for fluorescence signal output. On the basis of the amplification strategy, this biosensor displays excellent sensitivity and selectivity for ATP detection with an outstanding detection limit of 0.81 pM. Through simultaneously enhancing the target response signal value and reducing nonspecific background, this work deducted the background effect, and showed high sensitivity and reproducibility. Moreover, our biosensor also shows promising potential in real sample analysis. Therefore, the proximity-enabled BERA strategy indeed creates a simple and valuable fluorescence sensing platform for ATP identification and related disease diagnosis and biomedical research.

A supramolecule based fluorescence turn-on and ratiometric sensor for ATP in aqueous solution.

Considering the biological relevance of adenosine triphosphate (ATP) as an "energy currency" in all organisms and significance of its detection in various diseased conditions, enormous efforts have been made to develop selective and sensitive fluorescent sensors for the detection of ATP. However, these developed sensor probes frequently involve technically challenging and time-consuming synthetic protocols for the production of sensor molecules and often suffer from poor solubility in aqueous medium. Another major disadvantage of these developed sensor systems is their single wavelength based operation which makes their performance susceptible to minute changes in experimental conditions. Herein, we report a fluorescence turn-on ratiometric sensor for the detection of ATP which operates by the dissociation of Thioflavin-T-sulphated-β-cyclodextrin supramolecular assembly by Zn2+ followed by ATP induced reassociation of the same. This modulation of the monomer/aggregate equilibrium of the supramolecular assembly followed by subsequent interactions with Zn2+ and ATP acts as an optimal scheme for the ratiometric detection of ATP. Overall this supramolecular ensemble based sensing platform provides a simple, sensitive, selective and label free detection approach for ATP in aqueous solution. Importantly, our sensor platform responds to ATP in the biologically complex media of serum samples suggesting its potential for possible applications in real-life scenarios.

Adenosine triphosphate responsive metal-organic frameworks equipped with a DNA structure lock for construction of a ratiometric SERS biosensor.

A novel ratiometric surface-enhanced Raman scattering (SERS) biosensor was constructed based on stimuli-responsive DNA functionalized metal organic frameworks (MOFs) for detection of adenosine triphosphate (ATP). As a result, the detection range of ATP was 1 nM to 200 nM with a detection limit of 0.4 nM. The ratiometric SERS biosensor strategy offers a lower detection limit and exhibits a more enhanced performance than the typical SERS detection based on single signal response, which may have potential for detection of other biomolecules or metal ions.

A AuNP-capped cage fluorescent biosensor based on controlled-release and cyclic enzymatic amplification for ultrasensitive detection of ATP.

Gold nanodevices have attracted extensive interest in the detection of specific targets within cells. However, constructing gold sensing devices that can be activated by the simulation of remote applications remains a huge challenge. Here, we report a Au nanoparticle (AuNP)-capped cage fluorescent biosensor based on controlled-release and Exonuclease III (Exo III) assisted cyclic enzymatic amplification that can be activated by adenosine triphosphate (ATP). In the system, AuNPs were used as the building blocks to cap the pores of Au nanocages (AuNCs) loaded with Rhodamine B (RhB) molecules through the hybridization of DNA. The RhB fluorescent molecules were finally released with the help of Exo III in the presence of ATP for detection purposes. Ultimately, the biosensor leads to a wide linear ATP detection range from 1.0 × 10-9 to 1.0 × 10-7 M with a limit of detection (LOD) down to 0.88 nM. In addition, it also has good selectivity for ATP to distinguish between ATP and ATP analogues such as cytidine triphosphate (CTP), guanosine triphosphate (GTP), and uridine triphosphate (UTP). Therefore, as a convenient and sensitive biosensor, it is expected to be widely used in the biomedical field.

Ultrasensitive and label-free detection of ATP by using gold nanorods coupled with enzyme assisted target recycling amplification

Abnormal concentration of adenosine triphosphate (ATP) is directly asscociate with several diseases. Thus, sensitive detection of ATP is essential to early diagnosis of disease. Herein, we described an ultrasensitive strategy for ATP detection by using positively charged gold nanorods ((+)AuNRs) as an efficient fluorescence quenching platform, coupled with exonuclease Ⅲ (Exo Ⅲ) assisted target recycling amplification. To construct the sensor, DNA template that contained ATP aptamer was used for the formation of Ag nanoclusters signal probe (DNA/AgNCs), the structure of it could change to duplex after the interaction of it with ATP. Such DNA template or duplex DNA product could electrostatically adsorb onto (+)AuNRs surface, resulting in the quenching of the fluorescence signal due to the vicinity of AgNCs to (+)AuNRs. With the addition of Exo Ⅲ, DNA duplex could be hydrolyzed and released from (+)AuNRs surface, leading to the recovery of a strong fluorescent signal, while ATP could be regenerated for next target recycling. Combing the good fluorescence quenching ability of (+)AuNRs and the Exo Ⅲ assisted signal amplification, a low detection limit of 26 pM was achieved for ATP detection. Notably, the proposed method can be successfully applied for detecting ATP in serum samples, indicating a potential application value in early cancer diagnosis.

Coupling aptazyme and catalytic hairpin assembly for cascaded dual signal amplified electrochemiluminescence biosensing

Developing reliable and sensitive detection methods for adenosine triphosphate (ATP) is vital for both clinical diagnosis and food safety. In this work, by coupling aptazyme- and catalytic hairpin assembly (CHA)-based signal amplification and electrochemiluminescence (ECL), an ultrasensitive biosensor for sensing ATP was fabricated using Ru(bpy)32+-doped silica nanoparticles (RuSiO2) as ECL probes and a ferrocene-functionalized hairpin DNA (hairpin-Fc) as quencher. The aptazyme-triggered cleavage of the DNA substrate and the CHA reaction both led to the circular release of trigger DNA, resulting in a significant dual signal amplification, with unprecedented enhancement up to 940-fold. Moreover, the bioconjugation of the DNA substrate with Au@Fe3O4 facilitated the separation and purification of the released trigger DNA, and effectively reduced the background signal. As a result, the as-prepared ECL biosensor exhibited a much lower detection limit of 0.054 pM for ATP, compared to those in previous reports, and showed high reliability for ATP detection in both spiked serum samples and Staphylococcus aureus. This work offers a new perspective for designing nucleic acid-based signal amplification for detecting ATP in bacterial analysis and clinical diagnosis.

Highly Photoluminescent Carbon Dots Derived from Discarded Chewing Gum: toward Multiple Sensing of pH, Ferric Ion, and Adenosine Triphosphate

AbstractIn this paper, we develop a sustainable strategy to reuse discarded chewing gum for low‐cost synthesis of carbon dots (CDs) via a H2O2‐assisted hydrothermal method. The as‐prepared CDs have a large photoluminescence (PL) quantum yield (QY) of up to 25.7%, much higher than the most values of waste‐derived CDs. Interestingly, the PL intensity of CDs increases linearly as pH rises from 1 to 6, demonstrating the possibility of CDs for accurate pH monitoring in acidic conditions. It is suggested that the pH‐responsive PL feature is induced by the protonation and deprotonation of the surface oxygenous groups of CDs. Through the fluorescence quenching and recovery processes, the CDs can serve as a selective and sensitive sensor for sequential detection of ferric ion (Fe3+) and adenosine triphosphate (ATP) with limit of detection (LOD) as low as 0.028 and 0.003 μM, respectively. A possible electron transfer process is proposed to elucidate the sensing mechanism. For real applications, the CD sensing platform is successfully employed to determinate pH in natural stream, Fe3+ in tap water, and ATP in erythrocyte sample.

Design and Synthesis of Ag Nanocluster Molecular Beacon for Adenosine Triphosphate Detection.

This study presents a fluorescence method for detecting adenosine triphosphate (ATP) based on a label-free Ag nanocluster molecular beacon (MB) with high sensitivity. The sensor contains a hairpin-shaped MB, two short single-stranded DNA strands, and T4 DNA ligase. The MB consists of three parts, which are the template DNA sequence for synthesizing Ag nanoclusters at the 5′ end, the middle DNA with a hairpin-shaped structure, and the guanine base-rich DNA sequence at the 3′ end. The sensor exhibits high fluorescence intensity in the absence of ATP. However, when the probe is used for ATP detection, the two short DNA sequences in the sensor would form a long sequence by enzymatic ligation reaction; this long sequence opens the hairpin-shaped structure of the MB and decreases the fluorescence of the system. Under optimal analytical conditions, a clear linear relationship is observed between ATP concentration and fluorescence intensity in the range of 0.1–10 μM. The interference presented by other small molecules during ATP detection is evaluated, and results confirm the good selectivity of the proposed sensor. Compared with traditional methods, the sensor is label free, easy to operate, inexpensive, and highly sensitive.

Low field nuclear magnetic sensing technology based on hydrogel-coated superparamagnetic particles

Based on superparamagnetic nanoparticles, a responsive polyacrylamide hydrogel self-assembled by nucleic acid hairpin hybridization chain reaction was designed, and a universal low field nuclear magnetic resonance sensing platform was successfully constructed. As the target was gradually added, the hydrogel coating on the surface of the magnetic nanoparticle was opened layer by layer through binding with the aptamer, which specifically bonded thereto, causing different degrees of exposure of the magnetic nanoparticle, resulting in changes of low field nuclear magnetic resonance signals. This method was originally applied to the rapid detection of adenosine triphosphate (ATP), and the versatility of the method was verified using polychlorinated biphenyl 77 (PCB77). This method had the advantage of being fast, convenient, and low cost, and it can be easily operated with high repeatability. This universal method can detect a variety of targets by replacing aptamers and may be useful in controlling food quality and for rapidly detecting cancer cells in vitro.

ATP Mimics pH-Dependent Dual Peroxidase-Catalase Activities Driving H 2 O 2 Decomposition

Adenosine triphosphate (ATP) is produced mainly in the mitochondrion, and its primary task is to function as a ubiquitous energy currency to meet the cellular metabolic demands in biological system...

Transfer of mitochondria from mesenchymal stem cells derived from induced pluripotent stem cells attenuates hypoxia-ischemia-induced mitochondrial dysfunction in PC12 cells.

Mitochondrial dysfunction in neurons has been implicated in hypoxia-ischemia-induced brain injury. Although mesenchymal stem cell therapy has emerged as a novel treatment for this pathology, the mechanisms are not fully understood. To address this issue, we first co-cultured 1.5 × 105 PC12 cells with mesenchymal stem cells that were derived from induced pluripotent stem cells at a ratio of 1:1, and then intervened with cobalt chloride (CoCl2) for 24 hours. Reactive oxygen species in PC12 cells was measured by Mito-sox. Mitochondrial membrane potential (?Ψm) in PC12 cells was determined by JC-1 staining. Apoptosis of PC12 cells was detected by terminal deoxynucleotidal transferase-mediated dUTP nick end-labeling staining. Mitochondrial morphology in PC12 cells was examined by transmission electron microscopy. Transfer of mitochondria from the mesenchymal stem cells derived from induced pluripotent stem cells to damaged PC12 cells was measured by flow cytometry. Mesenchymal stem cells were induced from pluripotent stem cells by lentivirus infection containing green fluorescent protein in mitochondria. Then they were co-cultured with PC12 cells in Transwell chambers and treated with CoCl2 for 24 hours to detect adenosine triphosphate level in PC12 cells. CoCl2-induced PC12 cell damage was dose-dependent. Co-culture with mesenchymal stem cells significantly reduced apoptosis and restored ?Ψm in the injured PC12 cells under CoCl2 challenge. Co-culture with mesenchymal stem cells ameliorated mitochondrial swelling, the disappearance of cristae, and chromatin margination in the injured PC12 cells. After direct co-culture, mitochondrial transfer from the mesenchymal stem cells stem cells to PC12 cells was detected via formed tunneling nanotubes between these two types of cells. The transfer efficiency was greatly enhanced in the presence of CoCl2. More importantly, inhibition of tunneling nanotubes partially abrogated the beneficial effects of mesenchymal stem cells on CoCl2-induced PC12 cell injury. Mesenchymal stem cells reduced CoCl2-induced PC12 cell injury and these effects were in part due to efficacious mitochondrial transfer.

A Dual-Sensing DNA Nanostructure with an Ultrabroad Detection Range.

Despite considerable interest in the development of biosensors that can measure analyte concentrations with a dynamic range spanning many orders of magnitude, this goal has proven difficult to achieve. We describe here a modular biosensor architecture that integrates two different readout mechanisms into a single-molecule construct that can achieve target detection across an extraordinarily broad dynamic range. Our dual-mode readout DNA biosensor combines an aptamer and a DNAzyme to quantify adenosine triphosphate (ATP) with two different mechanisms, which respond to low (micromolar) and high (millimolar) concentrations by generating distinct readouts based on changes in fluorescence and absorbance, respectively. Importantly, we have also devised regulatory strategies to fine-tune the target detection range of each sensor module by controlling the target-sensitivity of each readout mechanism. Using this strategy, we report the detection of ATP at a dynamic range spanning 1-500 000 μM, more than 5 orders of magnitude, representing the largest dynamic range reported to date with a single biosensor construct.

Monitoring of dynamic ATP level changes by oligomycin-modulated ATP synthase inhibition in SW480 cancer cells using fluorescent \u201cOn-Off\u201d switching DNA aptamer

Adenosine triphosphate (ATP) is the main energy source in cells and an important biomolecule participating in cellular reactions in living organisms. Since the ATP level changes dynamically reflecting the development of a debilitating disease or carcinogenesis, we have focused in this work on monitoring of the oligomycin (OMC)-modulated ATP synthase inhibition using a fluorescent-switching DNA aptamer designed for the detection of ATP (Apt(ATP)), as the model for studies of dynamic ATP level variation. The behavior of the ATP aptamer has been characterized using fluorescence spectroscopy. The Intramolecular fluorescence resonance energy transfer (iFRET) operates in the proposed aptamer from the FAM dye moiety to guanines of the aptamer G-quadruplex when the target ATP is present and binds to the aptamer changing its conformation. The iFRET process enables the detection of ATP down to the limit of detection, LOD = 17 μM, without resorting to any extra chemi-amplification schemes. The selectivity coefficients for relevant interferent triphosphates (UTP, GTP, and CTP) are low for the same concentration as that of ATP. We have demonstrated an efficient transfection of intact cells and OMC-treated SW480 colon cancer cells with Apt(ATP), using microscopic imaging, iFRET measurements, and cell viability testing with MTT method. The applicability of the switching DNA aptamer for the analysis of real samples, obtained by lysis of SW480 cells, was also tested. The proposed Apt(ATP) may be considered as a viable candidate for utilization in measurements of dynamic ATP level modulation in cells in different stages of cancer development and testing of new drugs in pharmacological studies.Graphical abstract

Abundance and survival of microbial aerosols in the troposphere and stratosphere.

Bioaerosol transport in the atmosphere disperses microbial species between continents, affects human and plant health, and may influence hydrologic cycling. However, there have been few quantitative observations of bioaerosols at altitudes more than a few kilometers above the surface. Lack of data on bioaerosol distributions in the atmosphere has impeded efforts to assess the aerial dissemination of microbes and their vertical extent in the biosphere. In this study, a helium balloon payload system was used to sample microbial cells and dust particles in air masses as high as 38 km above sea level over three locations in the southwestern United States. The cell concentrations at altitudes between 3 and 29 km were highly similar (2-5 × 105 cells m-3) and approximately threefold lower than those observed in the convective boundary layer (CBL; 1 × 106 cells m-3), decreasing to 8 × 104 cells m-3 at 35-38 km. The detection of adenosine triphosphate (ATP) and recovery of bacteria possessing extreme tolerance to desiccation and shortwave ultraviolet radiation confirmed that certain microorganisms have the capacity to persist at lower altitudes of the stratosphere. Our data and related calculations provide constraints on the upper altitudinal boundary for microbial habitability in the biosphere.

Functionalized acupuncture needle as a SERS-active platform for rapid and sensitive determination of adenosine triphosphate.

The development of sensitive and rapid methods for analysis and detection of small molecules is highly desirable for medical diagnostics and therapeutics. We report an acupuncture needle functionalized with gold nanoparticles (Au NPs) and a macrocyclic amine (MA) Raman tag as the platform to realize the sensitive detection of adenosine triphosphate (ATP) by surface-enhanced Raman spectroscopy (SERS). The assembled Au NPs with abundant hot spots on the surface of the needle avoids the aggregation of Au NPs and results in a good signal response. Moreover, there is strong combination between ATP and MA through electrostatic adsorption, hydrogen-bonding interactions, and π-π stacking, and as a consequence, this functionalized needle can be used as a SERS platform for detection of ATP (25 nM) through a decrease of the Raman signal of MA resulting from the high chemical affinity of ATP for MA. Specially, the Au NP/MA-functionalized needle is conveniently used to monitor ATP (100 nM) added to serum, and demonstrates great promise in the study and detection of ATP in a complex sample, laying the foundation for SERS applications in complex acupuncture specimens with fast response and simple operation. Graphical abstract.

MIL/Aptamer as a Nanosensor Capable of Resisting Nonspecific Displacement for ATP Imaging in Living Cells.

Fluorescent probes physisorbed on nanomaterials have emerged as a kind of useful and facile sensing platform for biological important molecules. However, nonspecific displacement in the physisorption systems is a non-negligible problem for the intracellular analysis. MIL (Materials of Institut Lavoisier), a subclass of metal–organic frameworks (MOFs), has high porosity, large surface area, and intriguing three-dimensional (3D) nanostructure with promising biological and biomedical applications such as molecular detection and drug delivery. Herein, we report MIL/aptamer-FAM as a nanosensor capable of resisting nonspecific displacement for intracellular adenosinetriphosphate (ATP) sensing and imaging. In this approach, by virtue of the remarkable quenching capability, high affinity of aptamers, and dramatic capability of resisting nonspecific displacement of 3D MIL-100, the assay and imaging of ATP in living cells were realized. Our results demonstrated that the MIL/aptamer-FAM nanosensor not only shows high selectivity for the detection of ATP in buffer but also is able to act as a “signal-on” nanosensor for specific imaging of ATP in living cells. The strategy reported here opens up a new way to develop MOF-based nanosensors for intracellular delivery and metabolite detection.

A label-free fluorometric aptasensor for adenosine triphosphate (ATP) detection based on aggregation-induced emission probe

Based on Aggregation-Induced Emission (AIE), the development of a label-free, simple and sensitive fluorometric aptasensor for adenosine triphosphate (ATP) detection is described. With ATP present, the aptamers will combine with ATP and the conformation of the aptamer will switch from a random coil to an antiparallel G-quadruplex, which impedes the digestion by exonuclease I (Exo I). Addition of 4,4 -(1E,1E)-2,2-(anthracene-9,10-diyl) bis (ethene-2,1-diyl) bis (N,N, N-trimethyl-benzenaminium iodide) (DSAI) into the solution will cause aggregation of DSAI on the surface of the aptamer/ATP complex and consequently give rise to strong emission. Additionally, a good linear relationship was observed under optimized conditions between the fluorescence intensities and the logarithm of ATP concentrations (R2 = 0.9908). The established aptamer sensor was highly sensitive and exhibited a low limit of detection of 32.8 nM, with superior specificity for ATP. It was also used in the quantification of ATP levels in human serum samples and demonstrated satisfactory recoveries in the scope of 93.2%–107.6%. The cellular ATP assay results indicated that the developed method can be used for monitoring ATP concentrations in cell extracts without the interference of other substances in the cells. This method offers several advantages such as simplicity, rapidity, low cost and excellent selectivity, which make it hold great potential for the detection of ATP in bioanalytical and biological studies.

A highly sensitive fluorescence-enhanced aptasensor based on polyAn-aptamer nanostructure

According to the principle of metal-enhanced fluorescence (MEF), a localized surface plasmon resonance (LSPR) based method was used to develop a highly sensitive fluorescent aptasensor for detecting adenosine triphosphate (ATP). In this work, a polyAn-based special nucleic acid aptamers modified on gold nanoparticle (AuNPs) were hybridized with the FAM-labeled single-stand oligonucleotides. It was found that a great enhancement of fluorescence intensity in the nanostructures could be programmable regulated by adjusting the number of base A. The experiments showed the maximum enhancement for the fluorophore occurs when the polyAn length reached to 9 nm and, there was a good linear relationship between the fluorescence intensity and concentrations of ATP in the range of 0.5–1 nM. The detection limit obtained was 0.2 nM. Compared with the detection performances of traditional strategies, the sensitivity of the method was increased by more than 7 times, which make it hold great potential in bioanalytical and biological studies.

New Strategy for Ultrasensitive Aptasensor Fabrication: D-A-D Constitution as a Charge Transfer Platform and Recognition Element.

Over the past decade, various sensing systems based on aptamers have attracted a great deal of studies directed at designing highly selective biosensors. In this paper, we described a new-style electrochemical aptamer sensor (aptasensor) via a donor-acceptor link substrate, which was characterized by electrochemical methods and other helpful characterization instruments. Molecules with D-A-D configuration always undergo an intrinsic signal amplification due to the elongation of the π-electron conjugation. Triphenylamine, a peripheral electron donor, has excellent hole-transport property and is able to assemble on the surface of glassy carbon electrode by π-π stacking interaction. To further improve the performance of the adenosine triphosphate (ATP) sensor, we chose diphenylfumaronitrile-containing electron-withdrawing group as the central core to promote charge transfer, which can also efficiently combine with aptamers by multihydrogen bond function. Surprisingly, the sensing platform showed a wide liner range from 0.1 pM to 100 nM, with a detection limit of 0.018 pM. We examined the ATP in human serum sample, indicating that the novel aptasensor based on D-A-D conjugated polymer holds great possibility for practical detection of ATP. Moreover, it is foreseeable that the conjugated polymers of the D-A structure will have promising application in the preparation of biosensors.

Development of a point-of-contact technique to measure adenosine triphosphate: A quality improvement study.

PurposePatients with heart failure with preserved ejection fraction (HFpEF) experience fatigue due to impaired myocardial bioenergetics. Cardiomyocyte function depends on the delivery of adenosine triphosphate (ATP), yet there is no convenient bedside method to measure ATP. The purpose of this study was to develop a point-of-contact measurement of ATP that can be used in a clinical setting.MethodsIn a laboratory setting, digital finger punctures were conducted using 5 μl and 10 μl of capillary blood placed into various amounts of water (H2O). After mixing the solution for 10 s, a Hygiena AquaSnapTM Free ATP probe was placed into the solution for 10 s for the detection of ATP. The probe was then placed into the Hygiena luminometer for 15 s, and a value in relative light units (RLU) was obtained.ResultsTest samples using 10 μl of blood diluted from 50 to 500 mls of H2O produced ATP readings of 10,000-7569 RLUs. Using 5 μl of blood in 375–900 ml of H2O decreased the ATP values to 6459-4189 RLUs. Dilutional volume sparing experiments were conducted with ATP standards to determine the concentration of ATP per RLUs.ConclusionPatients with HFpEF have increased metabolic demand and impaired myocardial bioenergetics. Thus, identifying a method to measure ATP that is quick and accurate is imperative to accurately assess cellular energy production in this population. Point-of-contact measures, such as ATP, are needed for precision-guided treatment. Data from this study provides the first step toward developing evidence for health policies related to managing fatigue.