Bi-color FRET from two nano-donors to a single nano-acceptor: A universal aptasensing platform for simultaneous determination of dual targets
Graphical abstract
Introduction
Fluorescence resonance energy transfer (FRET) presents as a very promising tool for investigation of molecular interactions and molecular structure [1], [2], [3]. Two essential requirements for FRET occurrence are 1) a good spectral overlap for acceptor absorption and donor emission and 2) a close distance (<10 nm) between acceptor and donor [1]. Fortunately, separate donors and acceptors can be brought in close proximity through binding events such as protein interaction, antibody-based immunoreaction, and DNA hybridization [2], [4]. Disturbing the molecular interactions by the specific targets will lead to spatial separation of donor and acceptor probes which make donor–acceptor pair cannot engage in FRET process. As a result, FRET strategy becomes particular prevailing in biosensing owing to its target-specific signal and homogeneous assays without labor-intensive washing and separation steps [4]. In these designs, FRET assays can significantly improve the duration and simplicity of the experiment, in particular, can effectively avoid “false positive” results. Organic fluorophores are commonly served as either energy donor or energy acceptor in traditional FRET systems, for instance, Rhodamine B and methyl-red [5], Pyrene excimer-SYBR Green I [6], and Cy3-Cy5 [7] pairs. However, the optical limitations of organic fluorophores such as photobleaching, low resistance to chemicals, as well as short fluorescent lifetime, have seriously limited their broad biosensing applications [4].
Employing nanomaterials with appropriate optical features, to replace the traditional donor–acceptor pair, is certainly a central strategy to address the limitations of organic fluorophores. As a typical example, CdTe quantum dots (QDs)-Au nanoparticles (NPs), adopted as nano-donor and nano-acceptor pair, has aided advances in assembling biosensors for various targets ranged from small molecules [8], [9] to proteins [10], [11]. In most recent years, a cascade of FRET assays has been designed based on different nano-donor and nano-acceptor pairs like graphene QDs (GQDs)-AuNPs [12], GQDs-carbon nanotubes [13], carbon dots (CDs)-Au nanoclusters [14], [15], CDs-MnO2 nanostructures [16], GQDs-porphyrinic metal-organic frameworks [17], Au nanoclusters-MnO2 nanosheets [18], upconversion nanoparticles respectively paired to Ti3C2 nanosheets [19], Au nanorods [20], AuNPs [21] or ZnS NPs [22]. However, all these systems based on a single FRET pair are generally suitable for a single target detection in one measurement. Apparently, bi-color or multi-color FRET biosensors with an unambiguous separation of different nano-donor’s emission, if available, might provide a forceful technical support in advancing multiplexed analysis in a single measurement.
Mycotoxins, a group of toxic secondary metabolites generated by filamentous fungi, possess lethal threats to human or animals when they are appeared in food/feed [23], [24]. Aflatoxin B1 (AFB1), possessing the highest toxicity among various mycotoxins, has been defined as a group I carcinogen by the International Agency for Research on Cancer (IARC) in 1993 [25]. Ochratoxin A (OTA), listed as group IIB carcinogen by IARC, is another toxic mycotoxin which can be found in various foodstuffs [25]. What makes it worse is that the co-existence of AFB1 and OTA in a single sample leading to an enhanced toxicity owing to synergistic effect [26], [27]. Chromatographic methodologies including high-performance liquid chromatography (HPLC) [28] and HPLC or liquid chromatography coupled with tandem mass spectrometry [29], [30] are primarily used for multi-mycotoxin detection. However, these chromatographic methods suffer from expensive instruments, well-trained professionals and tedious procedures, which limit their applications in practice [31]. Thus, developing a sensitive, accurate, and rapid method for multi-mycotoxin detection is significant for food safety monitoring.
In this work, a simple-to-use aptasensor based on bi-color FRET from two nano-donors to a single nano-acceptor has been developed for simultaneous detection of AFB1 and OTA. Carbon dots (CDs) and CdZnTe QDs were regarded as two different nano-donors of FRET process, to label the respective aptamer against AFB1 and OTA. In another aspect, MoS2 nanosheets were selected as a single nano-acceptor paired to CDs and CdZnTe QDs. The van der Waals force between single-stranded aptamers and MoS2 nanosheets could bring the donor-acceptor pairs in near field and caused the energy transfer from donors to acceptor occur. By taking advantages of bi-color FRET from two nano-donors to a single nano-acceptor and target-induced fluorescence recovery, AFB1 and OTA can be simultaneously quantified in a single run.
Section snippets
Materials
We obtained AFB1, aflatoxin B2 (AFB2), fumonisin B1 (FB1), and ochratoxin A (OTA) from Sigma-Aldrich (USA). N-Hydroxysuccinimide (NHS), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), 3-mercaptopropionic acid (MPA), tris (hydroxymethyl) aminomethane (Tris), and other chemicals were obtained from Sinopharm Chemical Reagent Co., Ltd. (China). The aptamers possessing sequences of 5′-NH2-TGG GGT TTT GGT GGC GGG TGG TGT ACG GGC GAG GG-3′ (apt1, against AFB1) and 5′-NH2-GAT CGG
Characterization of MoS2 nanosheets
The as-prepared MoS2 nanosheets appeared to be black green in color (Inset a, Fig. 1A). The UV–vis absorption spectrum for MoS2 nanosheets suspension showed four characteristic absorption peaks, which located at 403, 450, 610, and 670 nm, respectively (Fig. 1A) [32], [36]. Two peaks at 610 and 670 nm can be respectively attributed to B and A excitonic peaks, resulting from the K point of the Brillouin zone in two-dimensional MoS2 nanosheet having large lateral dimensions [37]. The other two
Conclusions
In summary, we have designed a simple-to-use aptasensor for simultaneous detection of two mycotoxins on basis of the bi-color FRET from two nano-donors to a single nano-acceptor. In the detection diagram, two nano-donors (CDs and CdZnTe QDs) were used to label the aptamers specific for AFB1 and OTA, respectively. The single-stranded aptamers with two nano-donors could be flatly “lie” on MoS2 nanosheets surface through van der Waals force, which brought donor–acceptor in close proximity and
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgements
We thank for the financial support from National Natural Science Foundation of China (Nos. 21976071 and 21675066), Foundation of Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, Qingdao University of Science and Technology (No. SATM201807), Jiangsu University Fund (No. 19JDG025), and Cultivation Fund of Young Key Teacher at Jiangsu University (2015).
References (55)
- et al.
Recent applications of FRET-based multiplexed techniques
Trends Anal. Chem.
(2020) - et al.
FÖrster resonance energy transfer (FRET)-based biosensors for biological applications
Biosens. Bioelectron.
(2019) - et al.
Fluorescence resonance energy transfer based quantum dot-Aptasensor for the selective detection of microcystin-LR in eutrophic water
Chem. Eng. J.
(2019) - et al.
Graphene and graphene-like two-denominational materials based fluorescence resonance energy transfer (FRET) assays for biological applications
Biosens. Bioelectron.
(2017) - et al.
FRET-based fluorescent nanoprobe platform for sorting of active microorganisms by functional properties
Biosens. Bioelectron.
(2020) - et al.
Labeling-free fluorescent detection of DNA hybridization through FRET from pyrene excimer to DNA intercalator SYBR green I
Biosens. Bioelectron.
(2015) - et al.
Theoretical investigation on saturated Förster-resonant-energy-transfer microscopy using FRET dye pairs as fluorescent probes
Opt. Commun.
(2012) - et al.
Aptamer-based fluorescent detection of bisphenol A usingnonconjugated gold nanoparticles and CdTe quantum dots
Sensor. Actuat. B-Chem.
(2016) - et al.
A high-throughput homogeneous immunoassay based on Förster resonance energy transfer between quantum dots and gold nanoparticles
Anal. Chim. Acta
(2013) - et al.
A fluorescence resonance energy transfer (FRET) biosensor based on graphene quantum dots (GQDs) and gold nanoparticles (AuNPs) for the detection of mecA gene sequence of Staphylococcus aureus
Biosens. Bioelectron.
(2015)
DNA nanosensor based on biocompatible graphene quantum dots and carbon nanotubes
Biosens. Bioelectron.
Facile preparation of amino-carbon dots/gold nanoclusters FRET ratiometric fluorescent probe for sensing of Pb2+/Cu2+
Sensor. Actuat. B-Chem.
The construction of a FRET assembly by using gold nanoclusters and carbon dots and their application as a ratiometric probe for cysteine detection
Sensor. Actuat. B-Chem.
Carbon dot-MnO2 FRET system for fabrication of molecular logic gates
Sensor. Actuat. B-Chem.
FRET as a novel strategy to enhance the singlet oxygen generation of porphyrinic MOF decorated self-disinfecting fabrics
Chem. Eng. J.
DNA-templated Au nanoclusters and MnO2 sheets: a label-free and universal fluorescence biosensing platform
Sensor. Actuat. B-Chem.
Strand displacement dual amplification miRNAs strategy with FRET between NaYF4:Yb, Tm/Er upconversion nanoparticles and Ti3C2 nanosheets
Sensor. Actuat. B-Chem.
Aptamer-based sensing for thrombin in red region via fluorescence resonant energy transfer between NaYF4:Yb, Er upconversion nanoparticles and gold nanorods
Biosens. Bioelectron.
Upconversion nanoparticles based FRET aptasensor for rapid and ultrasenstive bacteria detection
Biosens. Bioelectron.
Monitoring and removal of trace heavy metal ions via fluorescence resonance energy transfer mechanism: In case of silver ions
Chem. Eng. J.
A fluorescence switch sensor used for D-Penicillamine sensing and logic gate based on the fluorescence recovery of carbon dots
Sensor. Actuat. B-Chem.
Target-driven switch-on fluorescence aptasensor for trace aflatoxin B1 determination based on highly fluorescent ternary CdZnTe quantum dots
Anal. Chim. Acta
Label-free fluorescence aptasensor for sensitive determination of bisphenol S by the salt-adjusted FRET between CQDs and MoS2
Sensor. Actuat. B-Chem.
Preparation and tribological properties of MoS2/graphene oxide composites
Appl. Surf. Sci.
Inkjet-printed electrochemically reduced graphene oxide microelectrode as a platform for HT-2 mycotoxin immunoenzymatic biosensing
Biosens. Bioelectron.
Preparation of graphene quantum dots based core-satellite hybrid spheres and their use as the ratiometric fluorescence probe for visual determination of mercury(II) ions
Anal. Chim. Acta
Aptamer induced multicoloured Au NCs-MoS2 “switch on” fluorescence resonance energy transfer biosensor for dual color simultaneous detection of multiple tumor markers by single wavelength excitation
Anal. Chim. Acta
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