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1.
Analyst ; 147(11): 2575-2581, 2022 May 30.
Article in English | MEDLINE | ID: mdl-35579472

ABSTRACT

In this paper, we developed an amplified fluorescence biosensor for acetylcholinesterase (AChE) activity detection by taking advantage of the mercury ion-mediated Mgzyme (Mg2+-dependent DNAzyme) activity. The catalytic activity of Mgzyme can be inhibited by the formation of T-Hg2+-T base pairs between the Mgzyme and mercury ions. Therefore, the Mgzyme-Hg2+ complex has no activity on a molecular beacon (MB) substrate, which afforded a very weak fluorescence background for this biosensor. After the addition of acetylcholinesterase (AChE), the substrate acetylthiocholine could be hydrolyzed to thiocholine, which has a stronger binding power with mercury ions than T-Hg2+-T base pairs. Therefore, the Mgzyme activity was recovered. The activated Mgzyme could hybridize with the MB substrate and undergo many cleavage cycles, resulting in a significant increase of fluorescence intensity. This biosensor displayed high sensitivity with the detection limit as low as 0.01 mU mL-1. Moreover, this design did not require complex composition and sequence design; thus it is simple and convenient. This biosensor was also applied for the determination of AChE in human blood and showed satisfactory results.


Subject(s)
Biosensing Techniques , DNA, Catalytic , Mercury , Acetylcholinesterase/metabolism , Biosensing Techniques/methods , DNA, Catalytic/chemistry , Humans , Ions , Limit of Detection , Mercury/chemistry
2.
ChemMedChem ; 16(16): 2426-2440, 2021 08 19.
Article in English | MEDLINE | ID: mdl-33780139

ABSTRACT

With the advantage of inherent responsiveness that can change the spectroscopic signals from "off" to "on" state in responding to targets (e. g. biological analytes/microenvironmental factors), activatable fluorescent probes have attracted extensive attention and made significant progress in the field of bioimaging and biosensing. Due to the high depth of tissue penetration, minimal tissue damage and negligible background signal at longer wavelengths, the development of second near-infrared window (NIR-II) fluorescent materials provides a new opportunity to develop activable fluorescent probes. Here, we summarized properties, advantages and disadvantages of mainly NIR-II fluorophores (such as rare earth-doped nanoparticles, quantum dots, single-walled carbon nanotubes, small molecule dyes, conjugated polymers and gold nanoclusters), then overviewed current role and development of activatable NIR-II fluorescent probes (AFPs) for biomedical applications including biosensing, bioimaging and therapeutic. The potential challenges and perspectives of AFPs in deep-tissue imaging and clinical application are also discussed.


Subject(s)
Biosensing Techniques , Fluorescent Dyes/chemistry , Optical Imaging , Animals , Drug Development , Fluorescent Dyes/chemical synthesis , Humans , Infrared Rays , Molecular Structure
3.
Analyst ; 144(6): 1982-1987, 2019 Mar 11.
Article in English | MEDLINE | ID: mdl-30694264

ABSTRACT

In this paper, by taking advantage of the fact that silver ions could mediate the Mg2+-dependent DNAzyme (Mgzyme) activity, we for the first time developed a turn-on fluorescent biosensor for amplified cysteine (Cys) detection. Because Mgzyme can interact with the silver ion and form cytosine-Ag+-cytosine (C-Ag+-C) base pairs, the conformation of its catalytic core was changed. As a result, the catalytic activity of Mgzyme was suppressed and the Mgzyme-Ag+ complex could not initiate the cleavage reaction. Therefore, the background fluorescence of the biosensor was very low. In the presence of Cys, Cys can bind tightly to the silver ion and disrupt the C-Ag+-C base pairs in the Mgzyme-Ag+ complex, leading to the restoration of Mgzyme activity. The activated Mgzyme could hybridize with the MB substrate and undergo many cleavage cycles, resulting in a significant increase of fluorescence intensity. This designed strategy provided amplified fluorescence detection of cysteine, with a detection limit of 2 nM. Moreover, the strong binding between Cys and Ag+ ensured that the biosensor had a desirable selectivity for Cys. This sensing system was also used to detect Cys in human urine samples and displayed satisfying results.


Subject(s)
Biosensing Techniques/methods , Cysteine/urine , DNA, Catalytic/metabolism , Fluorescence , Magnesium/chemistry , Silver/chemistry , DNA, Catalytic/chemistry , Humans , Limit of Detection
4.
Talanta ; 149: 98-102, 2016.
Article in English | MEDLINE | ID: mdl-26717819

ABSTRACT

In this work we use a water-soluble cationic perylene derivative (compound 1) as the G-quadruplex (G4) structure fluorescence indicator to construct a fluorescent biosensor for simple, rapid and label-free detection of Pb(2+). In the absence of Pb(2+), strong electrostatic interactions between compound 1 and the G-rich DNA probe (PW17) induced the aggregation of compound 1 and resulted in the fluorescence quenching. In the presence of Pb(2+), the PW17 formed Pb(2+)-stabilized G4 structure, which reduced the aggregation of compound 1 and gave rise to high fluorescence. This allowed us to use convenient "mix-and-detect" protocol for quantitative analysis of Pb(2+). Since Pb(2+) can specially induce PW17 to form compact DNA fold, our proposed biosensor displayed high selectivity for Pb(2+). It also exhibited a high sensitivity to Pb(2+), with a limit of detection of 5.0nM observed. Furthermore, the biosensor was applied for the detection of Pb(2+) in urine and paint samples, and both showed satisfactory results.


Subject(s)
Biosensing Techniques , Fluorescent Dyes/chemistry , G-Quadruplexes , Lead/analysis , Perylene/chemistry , DNA Probes , Fluorescence , Humans , Lead/chemistry , Limit of Detection , Paint/analysis , Urine/chemistry
5.
Chem Commun (Camb) ; 51(60): 12095-8, 2015 Aug 04.
Article in English | MEDLINE | ID: mdl-26120805

ABSTRACT

By employing DNAzyme as a recognition group and amplifier, and DNA-stabilized silver nanoclusters (DNA/AgNCs) as signal reporters, we reported for the first time a label-free catalytic and molecular beacon as an amplified biosensing platform for highly selective detection of cofactors such as Pb(2+) and L-histidine.


Subject(s)
Biosensing Techniques/methods , DNA, Catalytic/metabolism , DNA/metabolism , Histidine/analysis , Lead/analysis , Nanostructures/chemistry , Silver/chemistry , Cations, Divalent/analysis , Cations, Divalent/metabolism , DNA/chemistry , Histidine/metabolism , Lead/metabolism , Limit of Detection , Rivers/chemistry , Spectrometry, Fluorescence/methods
6.
Anal Chem ; 87(8): 4448-54, 2015 Apr 21.
Article in English | MEDLINE | ID: mdl-25791340

ABSTRACT

Many one-photon fluorescence-based theranostic nanosystems have been developed for simultaneous therapeutic intervention/monitoring for various types of cancers. However, for early diagnosis of cancer, two-photon fluorescence microscopy (TPFM) can realize deep-tissue imaging with higher spatial resolution. In this study, we first report a multiple functional nanoprobe for contrast-enhanced bimodal cellular imaging and targeted therapy. Components of the nanoprobe include (1) two-photon dye-doped mesoporous silica nanoparticles (TPD-MSNs); (2) MnO2 nanosheets that act as a (i) gatekeeper for TPD-MSNs, (ii) quencher for TP fluorescence, and (iii) contrast agent for MRI; (3) cancer cell-targeting aptamers. Guided by aptamers, TPD-MSNs are rapidly internalized into the target cells. Next, intracellular glutathione reduces MnO2 to Mn(2+) ions, resulting in contrast-enhanced TP fluorescence and magnetic resonance signal for cellular imaging. Meanwhile, preloaded doxorubicin and Chlorin e6 are released for chemotherapy and photodynamic therapy, respectively, with a synergistic effect and significantly enhanced therapeutic efficacy.


Subject(s)
Contrast Media , Molecular Imaging , Molecular Targeted Therapy , Nanoparticles/therapeutic use , Neoplasms/diagnosis , Neoplasms/drug therapy , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Contrast Media/chemistry , Fluorescence , Humans , Magnetic Resonance Imaging , Nanoparticles/chemistry , Photochemotherapy , Photosensitizing Agents/therapeutic use , Silicon Dioxide/chemistry , Tumor Cells, Cultured
7.
Chem Commun (Camb) ; 49(59): 6644-6, 2013 Jul 28.
Article in English | MEDLINE | ID: mdl-23771054

ABSTRACT

Taking advantage of the super-quenching effect of the cationic perylene derivative on adjacent fluorophores, we for the first time reported a DNAzyme-perylene complex-based strategy for constructing fluorescence catalytic biosensors with improved sensitivity.


Subject(s)
Biosensing Techniques , DNA, Catalytic/chemistry , Fluorescence , Perylene/chemistry , Catalysis , DNA, Catalytic/metabolism , Molecular Structure , Spectrometry, Fluorescence
8.
Anal Chem ; 85(7): 3614-20, 2013 Apr 02.
Article in English | MEDLINE | ID: mdl-23406194

ABSTRACT

DNAzymes have been widely applied as signal amplifiers for enzyme-free and highly sensitive detection of DNA. A few of them have also been employed for amplified detection of other biomolecules via a target-triggered assembly of split or mutated DNAzyme strategy. However, most of these designs adopt Mg(2+)-dependent DNAzyme as the catalytic unit, which suffered from low catalytic cleavage activity. Meanwhile, some DNAzymes with high catalytic activity are not suitable for these designs because the slight modification of the catalytic core might results in remarkably decreased or even no catalytic activity of these DNAzymes. On the basis of DNAzyme topological effect or the terminal protection of small-molecule-linked DNA, we developed two versatile sensing platforms for amplified detection of different biotargets. Since no modification is necessary for the catalytic core of the DNAzyme in these designs, they can employ any DNAzyme with high catalytic activity as amplified unit, which affords a high amplified efficiency for the sensing platform. A catalytic and molecular beacon design was further employed to realize the true enzymatic multiple turnover of DNAzyme. These designs together allow a high sensitivity for the biotargets, resulting in a detection limit of 20 pM, 0.2 U/mL, and 1 ng/mL for target DNA, DNA adenine methylation methyltransferase (Dam MTase), and streptavidin, respectively, much lower than previously reported biosensors. In addition, the proposed sensing strategy is versatile. By conjugating with various recognition units, it can be employed to detect a wide range of biotargets, varying from nucleic acids to proteins with high sensitivity.


Subject(s)
Biosensing Techniques/methods , DNA Modification Methylases/analysis , DNA, Catalytic/metabolism , DNA/analysis , Escherichia coli/enzymology , Proteins/analysis , Animals , Cell Line , DNA/metabolism , DNA Modification Methylases/metabolism , Enzyme Assays/methods , Humans , Limit of Detection , Proteins/metabolism
9.
Small ; 9(6): 951-7, 2013 Mar 25.
Article in English | MEDLINE | ID: mdl-23208981

ABSTRACT

Graphene nanomaterials are typically used in biosensing applications, and they have been demonstrated as good fluorescence quenchers. While many conventional amplification platforms are available, developing new nanomaterials and establishing simple, enzyme-free and low-cost strategies for high sensitivity biosensing is still challenging. Therefore, in this work, a core-shell magnetic graphitic nanocapsule (MGN) material is synthesized and its capabilities for the detection of biomolecules are investigated. MGN combines the unique properties of graphene and magnetic particles into one simple and sensitive biosensing platform, which quenches around 98% of the dye fluorescence within minutes. Based on a programmed multipurpose DNA capturing and releasing strategy, the MGN sensing platform demonstrates an outstanding capacity to fish, enrich, and detect DNA. Target DNA molecules as low as 50 pM could be detected, which is 3-fold lower than the limit of detection commonly achieved by carbon nanotube and graphene-based fluorescent biosensors. Moreover, the MGN platform exhibits good sensing specificity against DNA mismatch tests. Overall, therefore, these magnetic graphitic nanocapsules demonstrate a promising tool for molecular disease diagnosis and biomedicine. This simple fishing and enrichment strategy may also be extended to other biological and environmental applications and systems.


Subject(s)
DNA/analysis , Graphite/chemistry , Magnetics , Nanocapsules , Biosensing Techniques , Limit of Detection
10.
Anal Chim Acta ; 727: 67-70, 2012 May 21.
Article in English | MEDLINE | ID: mdl-22541825

ABSTRACT

Based on the super fluorescence quenching efficiency of graphene oxide and exonuclease III aided signal amplification, we develop a facile, sensitive, rapid and cost-effective method for DNA detection. In the presence of target DNA, the target-probe hybridization forms a double-stranded structure and exonuclease III catalyzes the stepwise removal of mononucleotides from the blunt 3' termini of probe, resulting in the recycling of the target DNA and signal amplification. Therefore, our proposed sensor exhibits a high sensitivity towards target DNA with a detection limit of 20 pM, which was even lower than previously reported GO-based DNA sensors without enzymatic amplification, and provides a universal sensing platform for sensitive detection of DNA.


Subject(s)
Biosensing Techniques/methods , DNA/analysis , Exodeoxyribonucleases/metabolism , Graphite/chemistry , Oxides/chemistry , Biocatalysis , Biosensing Techniques/instrumentation , DNA/metabolism , Electrochemical Techniques , Fluorescence , Spectrometry, Fluorescence
11.
Anal Chem ; 83(13): 5062-6, 2011 Jul 01.
Article in English | MEDLINE | ID: mdl-21639104

ABSTRACT

On the basis of the remarkable difference in affinity of graphene (GO) with ssDNA containing a different number of bases in length, we for the first time report a GO-DNAzyme based biosensor for amplified fluorescence "turn-on" detection of Pb(2+). A FAM-labeled DNAzyme-substrate hybrid acted as both a molecular recognition module and signal reporter and GO as a superquencher. By taking advantage of the super fluorescence quenching efficiency of GO, our proposed biosensor exhibits a high sensitivity toward the target with a detection limit of 300 pM for Pb(2+), which is lower than previously reported for catalytic beacons. Moreover, with the choice of a classic Pb(2+)-dependent GR-5 DNAzyme instead of 8-17 DNAzyme as the catalytic unit, the newly designed sensing system also shows an obviously improved selectivity than previously reported methods. Moreover, the sensing system was used for the determination of Pb(2+) in river water samples with satisfying results.


Subject(s)
Biosensing Techniques , DNA, Catalytic/chemistry , Graphite/chemistry , Lead/analysis , Fluorescence , Limit of Detection
12.
Anal Sci ; 26(5): 585-90, 2010.
Article in English | MEDLINE | ID: mdl-20467135

ABSTRACT

In this paper, we describe the fabrication and analytical characteristics of fluorescence-based copper ion-sensing glass slides. To construct the sensor, a naphthalimide derivative N-allyl-4-(bis(pyridin-2-ylmethyl)amino)ethylamino-1,8-naphthalimide (1) with a terminal double bond was synthesized and photo-copolymerized with 2-hydroxyethyl methacrylate (HEMA) on a glass surface treated with a silanizing agent. In the presence of Cu(2+) at pH 7.24, the resulting optical sensor undergoes fluorescence quenching. Thus, the proposed sensor with visible excitation can behave as a fluorescent sensor for the selective detection of Cu(2+). In addition, the sensor exhibits satisfactory selectivity, reproducibility and response time. The sensing membrane possesses a relatively long lifetime of at least 2 months. The linear response range covers a concentration range of Cu(2+) from 4.0 x 10(-7) to 6.0 x 10(-4) mol/L and the detection limit is 2.0 x 10(-7) mol/L. The determination of Cu(2+) in river water samples shows satisfactory results.


Subject(s)
Copper/analysis , Fluorescent Dyes/chemistry , Naphthalimides/chemistry , Water Pollutants, Chemical/analysis , Cations, Divalent/analysis , Cations, Divalent/chemistry , Copper/chemistry , Fluorescent Dyes/chemical synthesis , Glass/chemistry , Hydrogen-Ion Concentration , Limit of Detection , Methacrylates/chemistry , Naphthalimides/chemical synthesis , Photochemistry , Reproducibility of Results , Rivers/chemistry , Silanes/chemistry , Spectrometry, Fluorescence , Time Factors , Water Supply/analysis
13.
Anal Chim Acta ; 663(1): 85-90, 2010 Mar 17.
Article in English | MEDLINE | ID: mdl-20172101

ABSTRACT

A fluorescent probe 1 for Hg(2+) based on a rhodamine-coumarin conjugate was designed and synthesized. Probe 1 exhibits high sensitivity and selectivity for sensing Hg(2+), and about a 24-fold increase in fluorescence emission intensity is observed upon binding excess Hg(2+) in 50% water/ethanol buffered at pH 7.24. The fluorescence response to Hg(2+) is attributed to the 1:1 complex formation between probe 1 and Hg(2+), which has been utilized as the basis for the selective detection of Hg(2+). Besides, probe 1 was also found to show a reversible dual chromo- and fluorogenic response toward Hg(2+) likely due to the chelation-induced ring opening of rhodamine spirolactam. The analytical performance characteristics of the proposed Hg(2+)-sensitive probe were investigated. The linear response range covers a concentration range of Hg(2+) from 8.0x10(-8) to 1.0x10(-5)molL(-1) and the detection limit is 4.0x10(-8)molL(-1). The determination of Hg(2+) in both tap and river water samples displays satisfactory results.


Subject(s)
Coumarins/chemistry , Fluorescent Dyes/chemistry , Mercury/analysis , Organometallic Compounds/chemistry , Rhodamines/chemistry , Water Pollutants, Chemical/analysis , Fluorescent Dyes/chemical synthesis , Hydrogen-Ion Concentration , Organometallic Compounds/chemical synthesis
14.
Spectrochim Acta A Mol Biomol Spectrosc ; 73(4): 687-93, 2009 Aug 15.
Article in English | MEDLINE | ID: mdl-19398369

ABSTRACT

In the present paper, we describe the fabrication and analytical characteristics of fluorescence-based zinc ion-sensing glass slides. To construct the sensor, a benzoxazole derivative 4-benzoxazol-2'-yl-3-hydroxyphenyl allyl ether (1) with a terminal double bond was synthesized and copolymerized with 2-hydroxyethyl methacrylate (HEMA) on the activated surface of glass slides by UV irradiation. In the absence of Zn(2+) at pH 7.24, the resulting optical sensor emitted fluorescence at 450 nm via excited-state intramolecular proton transfer (ESIPT). Upon binding with Zn(2+), the ESIPT process was inhibited resulting in a 46 nm blue-shift of fluorescence emission. Thus, the proposed sensor can behave as a ratiometric fluorescent sensor for the selective detection of Zn(2+). In addition, the sensor shows nice selectivity, good reproducibility and fast response time. Cd(2+) did not interfere with Zn(2+) sensing. The sensing membrane demonstrates a good stability with a lifetime of at least 3 months. The linear response range covers a concentration range of Zn(2+) from 8.0x10(-5) to 4.0x10(-3) mol/L and the detection limit is 4.0x10(-5) mol/L. The determination of Zn(2+) in both tap and river water samples shows satisfactory results.


Subject(s)
Benzoxazoles/chemistry , Cations/analysis , Environmental Monitoring/instrumentation , Spectrometry, Fluorescence/methods , Water/analysis , Zinc/analysis , Benzoxazoles/chemical synthesis , Environmental Monitoring/economics , Environmental Monitoring/methods , Hydrogen-Ion Concentration , Reproducibility of Results , Rivers/chemistry , Sensitivity and Specificity , Time Factors , Water Pollutants, Chemical/analysis
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