ABSTRACT
Biomarkers detection in blood with high accuracy is crucial for the diagnosis and treatment of many diseases. In this study, the proof-of-concept fabrication of a dual-mode sensor based on a single probe (Re-BChE) using a dual-signaling electrochemical ratiometric strategy and a "turn-on" fluorescent method is presented. The probe Re-BChE was synthesized in a single step and demonstrated dual mode response toward butyrylcholinesterase (BChE), a promising biomarker of Alzheimer's disease (AD). Due to the specific hydrolysis reaction, the probe Re-BChE demonstrated a turn-on current response for BChE at -0.28 V, followed by a turn-off current response at -0.18 V, while the fluorescence spectrum demonstrated a turn-on response with an emission wavelength of 600 nm. The developed ratiometric electrochemical sensor and fluorescence detection demonstrated high sensitivity with BChE concentrations with a low detection limit of 0.08 µg mL-1 and 0.05 µg mL-1, respectively. Importantly, the dual-mode sensor presents the following advantages: (1) dual-mode readout can correct the impact of systematic or background error, thereby achieving more accurate results; (2) the responses of dual-mode readout originate from two distinct mechanisms and relatively independent signal transduction, in which there is no interference between two signaling routes. Additionally, compared with the reported single-signal electrochemical assays for BChE, both redox potential signals were detected in the absence of biological interference within a negative potential window. Furthermore, it was discovered that the outcomes of direct dual-mode electrochemical and fluorescence quantifications of the level of BChE in serum were in agreement with those obtained from the use of commercially available assay kits for BChE sensing. This method has the potential to serve as a useful point-of-care tool for the early detection of AD.
Subject(s)
Alzheimer Disease , Butyrylcholinesterase , Humans , Alzheimer Disease/diagnosis , Fluorescent Dyes , BiomarkersABSTRACT
Beta-site secretase (BACE1) catalyzes the cleavage of amyloid precursor protein (APP), which process ultimately lead to plaque deposition in the brain of Alzheimer's disease (AD). Thus, accurate monitor of BACE1 activity is essential to screen inhibitors for AD treatment. This study develops a sensitive electrochemical assay for probing BACE1 activity based on silver nanoparticles (AgNPs) and tyrosine conjugation as tags and a marking method, respectively. An APP segment is firstly immobilized on aminated microplate reactor. Cytosine (C) rich sequence-templated AgNPs/Zr-based metal-organic framework (MOF) composite is modified by phenol groups, and then the prepared tag (ph-AgNPs@MOF) is captured in microplate surface by the conjugation reaction of phenolic groups between tyrosine and tag. After cleavage by BACE1, the solution containing ph-AgNPs@MOF tags is transferred to the screen-printed graphene electrode (SPGE) surface for voltammetric detection of AgNP signal. This sensitive detection for BACE1 provided an excellent linear relationship between 1 to 200â pM with a detection limit of 0.8â pM. Furthermore, this electrochemical assay is successfully applied for screening of BACE1 inhibitors. This strategy is also verified to be used for evaluation of BACE1 in serum samples.
Subject(s)
Alzheimer Disease , Metal Nanoparticles , Metal-Organic Frameworks , Humans , Amyloid Precursor Protein Secretases/metabolism , Amyloid beta-Peptides/metabolism , Aspartic Acid Endopeptidases , Silver , Tyrosine , Alzheimer Disease/metabolism , Amyloid beta-Protein Precursor/metabolismABSTRACT
This study developed a novel luminescent assay for kinase activity using metal-organic coordination polymer nanoparticles (Tb/ATP-Zn) as the probe. Tb/ATP-Zn, self-assembled by adenosine triphosphate (ATP), Zn2+, and Tb3+, is non-luminescent. Protein kinase A (PKA) can catalyze the transformation of ATP within Tb/ATP-Zn nanoparticles to adenosine diphosphate (ADP), which in turn effectively sensitizes the luminescence of Tb3+. Based on this mechanism, Tb/ATP-Zn can realize the facile luminescent "turn-on" sensing of protein kinase activity without the use of external ATP and substrate peptide. Under optimized conditions, the fluorescence intensities of Tb/ATP-Zn at 550 nm are linear with the PKA activity within a range of 0.3-1.5 U·µL-1. The LOD (S/N = 3) of this method is down to 0.001 U·µL-1. The presented assay also features high selectivity, long-term stability, fast response, and convenient operation. Furthermore, Tb/ATP-Zn was successfully employed for monitoring PKA activity in cell lysis solutions. Probe Tb/ATP-Zn is thus expectable to be a powerful tool for the practical study of PKA in relevant biological events.
Subject(s)
Lanthanoid Series Elements , Metal Nanoparticles , Adenosine Triphosphate , Luminescence , Luminescent Measurements/methods , Polymers , Protein KinasesABSTRACT
Zinc, which is the second most abundant trace element in the human central nervous system, is closely associated with Alzheimer's disease (AD). However, attempts to develop highly sensitive and selective sensing systems for Zn2+ in the brain have not been successful. Here, we used a one-step solvothermal method to design and prepare a metal-organic framework (MOF) containing the dual ligands, terephthalic acid (H2BDC) and 2,2':6',2â³-terpyridine (TPY), with Eu3+ as a metal node. This MOF is denoted as Eu-MOF/BDC-TPY. Adjustment of the size and morphology of Eu-MOF/BDC-TPY allowed the dual ligands to produce multiple luminescence peaks, which could be interpreted via ratiometric fluorescence to detect Zn2+ using the ratio of Eu3+-based emission, as the internal reference, and ligand-based emission, as the indicator. Thus, Eu-MOF/BDC-TPY not only displayed higher selectivity than other metal cations but also offered a highly accurate, sensitive, wide linear, color change-based technique for detecting Zn2+ at concentrations ranging from 1 nM to 2 µM, with a low limit of detection (0.08 nM). Moreover, Eu-MOF/BDC-TPY maintained structural stability and displayed a fluorescence intensity of at least 95.4% following storage in water for 6 months. More importantly, Eu-MOF/BDC-TPY sensed the presence of Zn2+ markedly rapidly (within 5 s), which was very useful in practical application. Furthermore, the results of our ratiometric luminescent method-based analysis of Zn2+ in AD mouse brains were consistent with those obtained using inductively coupled plasma mass spectrometry.
Subject(s)
Alzheimer Disease , Lanthanoid Series Elements , Metal-Organic Frameworks , Alzheimer Disease/diagnosis , Animals , Europium/chemistry , Humans , Ligands , Luminescence , Metal-Organic Frameworks/chemistry , Mice , Microdialysis , ZincABSTRACT
As a vital gaseous signal molecule involved in various physiological and pathological processes, nitric oxide (NO) has attracted extensive attention in the last few decades. In this work, a copper and cobalt element-doped, biphenyl-(3, 4', 5)-tricarboxylic acid (H3PTC)-synthesized metal organic framework (CuCo-PTC MOF) composite with catalytic ability was synthesized by solvothermal method. The material can catalyse the oxidation of o-phenylenediamine (OPD) groups by hydrogen peroxide (H2O2) to form fluorophores (OPDox) with yellow fluorescence emission and greatly improves its reaction rate. In the presence of NO, OPD will react with NO to produce N-(2-hydrazinophenyl) methylamine, and the group will not react with H2O2. Therefore, the concentration of NO can be measured indirectly by comparing the changes of fluorescence intensity in the presence and absence of NO. As the concentration of NO changes, the change of solution colour (from bright yellow to colourless) can also be observed under a 365-nm UV lamp. Furthermore, the method represents high selectivity for NO and shows a fast (within 5 min) and specific fluorescence response toward NO with a linear range from 0.25 to 2.0 µM; the strategy has a limit of detection (LOD) of 0.15 µM. More importantly, the probe was successfully used to detect NO in cell lysate. The recovery was between 98.5 and 103.6%, and the relative standard deviation was between 0.4 and 1.8%. The endogenous NO in cells was successfully detected under the stimulation of L-arginine, which proved the possibility of the probe in real-time and rapid sensing in actual samples and cells. The results indicate that this sensing strategy has the potential to detect NO in the internal environment. Schematic of fluorescence detection of NO.
Subject(s)
Metal-Organic Frameworks , Catalysis , Hydrogen Peroxide , Limit of Detection , Nitric OxideABSTRACT
R-CDAs have been synthesized in a one-pot solvothermal procedure starting from 3,4-diaminobenzoic acid in an acidic medium. Transmission electron microscopy (TEM) revealed that R-CDAs nanoparticles exhibited a much larger diameter of 7.2-28.8 nm than traditional monodisperse carbon dots. X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FT-IR) revealed the presence of polar functional groups (hydroxyl, amino, carboxyl) on the surface of R-CDAs. Upon excitation with visible light (550 nm), R-CDAs emit stable, red fluorescence with a maximum at 610 nm. Under the optimum conditions, Cu2+ ions quench the fluorescence of this probe, and the signal is linear in a concentration range of copper ions between 5 and 600 nM with the detection limit of only 0.4 nM. Recoveries from 98.0 to 105.0% and relative standard deviations (RSD) from 2.8 to 4.5% have been obtained for detection of Cu2+ in real water samples. Furthermore, the R-CDAs fluorescent probe showed negligible cytotoxicity toward HeLa cells and good bioimaging ability, suggesting its potential applicability as a diagnostic tool in biomedicine.
Subject(s)
Carbon , Fluorescent Dyes , Humans , Fluorescent Dyes/toxicity , Carbon/toxicity , HeLa Cells , Spectroscopy, Fourier Transform InfraredABSTRACT
Brain pH has been proven to be a key factor in maintaining normal brain function. The relationship between local pH fluctuation and brain disease has not been extensively studied due to lack of the accurate in situ analysis technology. Herein, we have for the first time proposed a voltammetric pH sensor by measuring the ratio of current signals instead of the previously reported potential based on the Nernst equation. Single-walled carbon nanotubes (CNT) were first self-assembled on the electrode surface of a carbon-fiber nanotip electrode (CFNE). Then, poly-o-phenylenediamine (PoPD) molecules were deposited as pH-responsive molecules through in situ electrochemical polymerization. The compact CFNE/CNT/PoPD exhibited a good redox process with the on-off-on ratiometric electrochemical response to pH ranging from 4.5 to 8.2, providing self-correction for in situ pH detection. Thus, the proposed sensor enabled the accurate measurement of pH with excellent selectivity even in the presence of proteins or electroactive species. In addition, the sensor showed high repeatability, reproducibility, and reversibility in measuring pH and even demonstrated good stability when it was exposed to air for 5 months. Finally, we successfully detected the fluctuation of pH in rat brains with cerebral ischemia and rat whole blood. Overall, this research not only provides a good tool for the detection of rat brain pH but also provides a new strategy for further designing nanosensors for intracellular or subcellular pH.
Subject(s)
Nanotubes, Carbon , Animals , Brain , Electrochemical Techniques , Electrodes , Hydrogen-Ion Concentration , Rats , Reproducibility of ResultsABSTRACT
Redox homeostasis between hypochlorous acid (HClO/ClO-) and ascorbic acid (AA) significantly impacts many physiological and pathological processes. Herein, we report a new electrochemical sensor for the simultaneous determination of HClO and AA in body fluids. We first coated a carbon fiber microelectrode (CFME) with a three-dimensional nanocomposite consisting of graphene oxide (GO) and carbon nanotubes (CNTs) to fabricate the CFME/GO-CNT electrode. After the electrochemical reduction of GO (ERGO), we integrated a latent 1-(3,7-bis(dimethylamino)-10H-phenothiazin-10-yl)-2-methylpropan-1-one (MBS) electrochemical molecular recognition probe to monitor HClO and employed anthraquinone (AQ) as an internal reference. The compact CFME/ERGO-CNT/AQ + MBS sensor enabled the accurate and simultaneous measurement of HClO and AA with excellent selectivity and sensitivity. Measurements were highly reproducible, and the sensor was stable and exceptionally biocompatible. We successfully detected changes in the redox cycles of HClO and AA in human body fluids. This sensor is a significant advance for the investigation of reactions involved in cellular redox regulation. More importantly, we have devised a strategy for the design and construction of ratiometric electrochemical biosensors for the simultaneous determination of various bioactive species.
Subject(s)
Ascorbic Acid/analysis , Body Fluids/chemistry , Electrochemistry/instrumentation , Hypochlorous Acid/analysis , Ascorbic Acid/chemistry , Carbon Fiber/chemistry , Graphite/chemistry , Humans , Hypochlorous Acid/chemistry , Limit of Detection , Oxidation-Reduction , Time FactorsABSTRACT
A novel magnetic organic porous polymer (denoted as Fe3O4@PC-POP) was developed for magnetic solid-phase extraction (MSPE) of two gastric cancer biomarkers (P-cresol and 4-hydroxybenzoic acid) from urine samples prior to high-performance liquid chromatographic analysis. The adsorbent was characterized by scanning electron microscope, transmission electron microscope, FTIR, powder X-ray diffraction, and other techniques. The result of dynamic light scattering shows that the particle size of the adsorbent is mainly distributed around 400 nm. Based on the design concept of the Fe3O4@PC-POP, the proposed material can effectively capture the target analytes through electrostatic and hydrophobic interaction mechanism. Furthermore, the enrichment conditions were optimized by the response surface method, and the method was utilized for the determination of P-cresol and 4-hydroxybenzoic acid in real urine samples from health and gastric cancer patients with high enrichment factors (34.8 times for P-cresol and 38.7 times for 4-hydroxybenzoic acid), low limit of detection (0.9-5.0 µg L-1), wide linear ranges (3.0-1000 µg L-1), satisfactory relative standard deviation (2.5%-8.5%), and apparent recoveries (85.3-112% for healthy people's and 86.0-112% for gastric cancer patients' urine samples). This study provides a guided principle for design of the versatile polymer with specific capturing of the target compounds from complex biological samples. Graphical abstract.
Subject(s)
Biomarkers, Tumor/urine , Cresols/urine , Magnetite Nanoparticles/chemistry , Parabens/analysis , Polymers/chemistry , Stomach Neoplasms/urine , Adsorption , Biomarkers, Tumor/chemistry , Biomarkers, Tumor/isolation & purification , Chromatography, High Pressure Liquid , Cresols/chemistry , Cresols/isolation & purification , Humans , Limit of Detection , Parabens/chemistry , Parabens/isolation & purification , Piperazines/chemistry , Porosity , Solid Phase Extraction/methodsABSTRACT
A strategy for amplifying the signal of surface plasmon resonance (SPR) biosensors is reported. Biotinylated phenylalanine (Biotin-Phe) monomers were rapidly self-assembled into nanoparticles in a mild environment. The self-assembled nanoparticles were then used as the carriers of streptavidin-antibody complexes by the streptavidin-biotin interaction. The signal was amplified because of the high molecular weight of the nanoparticle-streptavidin-antibody conjugate. With prostate-specific antigen as a model analyte, the target concentration as low as 1 pg mL-1 was readily measured. The results of the nanoparticle-enhanced SPR biosensor for analysis of serum samples are well consistent with those achieved by the enzyme-linked immunosorbent assays. This work is valuable for designing of various optical and electronic biosensors through the streptavidin-biotin interaction. Graphical abstract.
Subject(s)
Biotin/analogs & derivatives , Nanoparticles/chemistry , Phenylalanine/analogs & derivatives , Surface Plasmon Resonance/methods , Antibodies, Immobilized/immunology , Humans , Limit of Detection , Male , Prostate-Specific Antigen/blood , Prostate-Specific Antigen/immunology , Streptavidin/chemistryABSTRACT
Thiophenol (PhSH) belongs to a class of highly reactive and toxic aromatic thiols with widespread applications in the chemical industry for preparing pesticides, polymers, and pharmaceuticals. In this review, we comprehensively summarize recent progress in the development of fluorescent probes for detecting and imaging PhSH. These probes are classified according to recognition moieties and are detailed on the basis of their structures and sensing performances. In addition, prospects for future research are also discussed.
Subject(s)
Fluorescent Dyes/chemistry , Phenols/chemistry , Sulfhydryl Compounds/chemistry , Amides/chemistry , Humans , Structure-Activity RelationshipABSTRACT
Hydrazine (N2 H4 ) is an important and commonly used chemical reagent for the preparation of textile dyes, pharmaceuticals, pesticides and so on. Despite its widespread industrial applications, hydrazine is highly toxic and exposure to this chemical can cause many symptoms and severe damage to the liver, kidneys, and central nervous system. As a consequence, many efforts have been devoted to the development of fluorescent probes for the selective sensing and/or imaging of N2 H4 . Although great efforts have been devoted in this area, the large number of important recent studies have not yet been systematically discussed in a review format so far. In this review, we have summarized the recently reported fluorescent N2 H4 probes, which are classified into several categories on the basis of the recognition moieties. Moreover, the sensing mechanism and probes designing strategy are also comprehensively discussed on aspects of the unique chemical characteristics of N2 H4 and the structures and spectral properties of fluorophores.
Subject(s)
Fluorescent Dyes/chemistry , Hydrazines/analysis , Hydrazines/chemistry , Hydrogen Bonding , Ketones/chemistry , Levulinic Acids/chemistry , Nitriles/chemistryABSTRACT
In this work, a lateral moving multi-pedal DNA tetrahedron machine (MTM) is designed and coupled with dual-signal output system to construct a biomimetic electrochemical ratiometric strategy for ultrasensitive target DNA analysis. The tetrahedral structure provided rigid support for the pedal, ensuring efficient replacement of the rail chain modified with ferrocene. By conjugating cholesterol molecules to one vertex of MTM, it is decorated on a lipid bilayer. This molecular architecture confers lateral movement of MTM on an electrode surface while prevents its detachment from the system. The methylene blue tagged hairpin probe provides constant power to support MTM swim on lipid bilayer. Compared with the conventional motion mode, the lateral moving mechanism has the fastest reaction rate and the highest signal-to-noise ratio. Additionally, the dual-signal reporting system further improves the accuracy of target detection on the basis of ensuring motion efficiency. The work improved movement efficiency and shortened time fragment. A linear relationship between the ratio value of two reporters and target DNA concentration was observed from 0.5 fM to 50 pM with a detection limit of 28 aM. The lateral motion mode of DNA machine coalescing with ratiometric system made this sensing platform ultrasensitive and accurate, which holds new avenue of early diagnosis.
Subject(s)
Biomimetics , Biosensing Techniques , Lipid Bilayers , Electrochemical Techniques , Limit of Detection , DNA/chemistryABSTRACT
Silver-doped hollow carbon spheres (Ag@HCS) were firstly introduced as electrochemical probes for glycated hemoglobin (HbA1c) sensing at a molecularly imprinted polymer (MIP)-based carbon cloth (CC) electrode. Herein, Ag@HCS was prepared using one-pot polymerization of resorcinol and formaldehyde with AgNO3 on the SiO2 template, subsequent carbonization, and template removal. Furthermore, poly-aminophenylboronic acid (PABA) as the MIP film was used as a sensing platform for recognition of HbA1c, which captured the Ag@HCS probe by binding of HbA1c with aptamer modified on the probe surface. Due to regular geometry, large specific surface area, superior electrical conductivity, and highly-dispersed Ag, the prepared Ag@HCS probe provided an amplified electrochemical signal based on the Ag oxidation. By use of the sandwich-type electrochemical sensor, the ultrahigh sensitivity of 4.365 µA (µg mL-1)-1 cm-2 and a wide detection range of 0.8-78.4 µg mL-1 for HbA1c detection with a low detection limit of 0.35 µg mL-1 were obtained. Excellent selectivity was obtained due to the specific binding between HbA1c and PABA-based MIP film. The fabricated electrochemical sensing platform was also implemented successfully for the determination of HbA1c concentrations in the serum of healthy individuals.
Subject(s)
Carbon , Molecular Imprinting , Humans , Silver , Glycated Hemoglobin , Silicon Dioxide , Electrochemical Techniques , 4-Aminobenzoic Acid , Polymers , Molecularly Imprinted Polymers , Limit of Detection , ElectrodesABSTRACT
Detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV CoV-2) pathogen and protein biomarkers can improve the diagnosis accuracy for Coronavirus disease 2019 (COVID-19). Electrochemical biosensors have attracted extensive attention in the scientific community because of their simple design, fast response, good portability, high sensitivity and high selectivity. In this review, we summarized the progress in the electrochemical detection of COVID-19 pathogen and SARS-CoV-2 biomarkers, including SARS-CoV-2 spike protein and nucleocapsid protein and their antibodies.
ABSTRACT
Insulin, a peptide hormone secreted by pancreatic ß cells, affects the development of diabetes and associated complications. Herein, we propose an electrochemical aptasensor for sensitive and selective detection of insulin using laser-scribed graphene electrodes (LSGEs). Before using disposable LSGEs, the development and proof-of-concept sensing experiments were firstly carried out on research-grade glassy carbon electrode (GCE). The aptasensor is based on using Exonuclease I (Exo I) that catalyses the hydrolysis of single-stranded aptamers attached to the electrode surface; however, the hydrolysis does not occur if the insulin is bound to the aptamer. Therefore, the unbound aptamers are cleaved by Exo I while insulin-bound aptamers remain on the electrode surface. In the next step, the gold nanoparticle - aptamer (AuNPs-Apt) probes are introduced to the electrode surface to form a 'sandwich' structure with the insulin on the surface-attached aptamer. The redox probe, methylene blue (MB), intercalates into the aptamers' guanine bases and the sandwich structure of AuNPs-Apt/insulin/surface-bound aptamer amplifies electrochemical signal from MBs. The signal can be well-correlated to the concentrations of insulin. A limit of detection of 22.7 fM was found for the LSGE-based sensors and 9.8 fM for GCE-based sensors used for comparison and initial sensor development. The results demonstrate successful fabrication of the single-use and sensitive LSGEs-based sensors for insulin detection.
Subject(s)
GraphiteABSTRACT
Alzheimer's disease (AD) is a degenerative disease of the central nervous system. The pathogenesis is still not fully clear. One of the main histopathological manifestations is senile plaques formed by ß-amyloid (Aß) accumulation. Aß is generated from the sequential proteolysis of amyloid precursor protein (APP) by ß-secretase [i.e. ß-site APP cleaving enzyme 1 (BACE1)] and γ-secretase, with a rate-limiting step controlled by BACE1 activity. Therefore, inhibiting BACE1 activity has become a potential therapeutic strategy for AD. The development of reliable detection methods for BACE1 activity plays an important role in early diagnosis of AD and evaluation of the therapeutic effect of new drugs for AD. This article has reviewed the recent advances in BACE1 activity detection techniques. The challenges of applying these analysis techniques to early clinical diagnosis of AD and development trends of the detection techniques have been prospected.
ABSTRACT
Rapid, simple, accurate and highly sensitive detection of enzymes is essential for early screening and clinical diagnosis of many diseases. In this study, we report the fabrication of a turn-on ratiometric electrochemical sensor for the in situ determination of ß-Galactosidase (ß-Gal) based on surface engineering and the design of a molecular probe (Pygal) specific for ß-Gal recognition. First, Pygal probe was synthesized and characterized, and then co-assembled with the methylene blue (MB) internal reference probe on the surface of single-wall carbon nanotubes (SWCNT)-modified carbon fiber microelectrode (CFME). The resulting CFME/SWCNT/MB + Pygal sensor is activated in the presence of ß-Gal giving one peak at 0.33 V originating from the oxidation of the product of Pygal enzymatic hydrolysis (PyOH). Another oxidation peak attributed to MB appears simultaneously at -0.28 V allowing the construction of a ratiometric electrochemical sensor for ß-Gal detection with improved sensitivity and accuracy. The sensor showed a linear response to ß-Gal in a wide concentration range from 1.5 to 30 U L-1 and a low detection limit of 0.1 U L-1. Moreover, the sensor demonstrated excellent selectivity against several biologically relevant hydrolases and redox-active molecules. Finally, the combination of excellent electrochemical performance and favorable physicochemical properties of CFME allowed the determination of ß-Gal in the whole blood of Parkinson's Disease (PD) model mice. The workflow reported in this study provides a strategy for the design and development of sensors for the in vivo monitoring of other enzymes important for the early diagnosis of different health issues.
Subject(s)
Biosensing Techniques , Nanotubes, Carbon , Parkinson Disease , Animals , Biosensing Techniques/methods , Carbon Fiber , Electrochemical Techniques/methods , Limit of Detection , Methylene Blue/chemistry , Mice , Parkinson Disease/diagnosis , beta-GalactosidaseABSTRACT
Small-molecule photosensitizers have great application prospects in photoelectrochemical (PEC) sensing due to their defined composition, diversified structure, and adjustable photophysical properties. Herein, we propose a new strategy for PEC analysis based on the target-induced in situ formation of the organic photosensitizer. Taking thiophenol (PhSH) as a model analyte, we designed and synthesized a 2,4-dinitrophenyl (DNP)-caged coumarin precursor (Dye-PhSH), which was then covalently coupled onto the TiO2 nanoarray substrate to obtain the working photoanode. Due to the intramolecular photoinduced electron transfer process, Dye-PhSH has only a very weak photoelectric response. Upon reacting with the target, Dye-PhSH undergoes a tandem reaction of the detachment of the DNP moiety and the intramolecular cyclization process, which leads to a coumarin dye with a pronounced photoelectric effect, thus achieving a highly selective turn-on PEC response to PhSH. For the first time, this study was to construct a PEC sensor by exploiting specific organic reactions for the in situ generation of small molecule-based photoactive material. It can be anticipated that the proposed strategy will expand the paradigm of PEC sensing and holds great potential for detecting various other analytes.
Subject(s)
Biosensing Techniques , Electrochemical Techniques , Photosensitizing AgentsABSTRACT
Interferon-gamma (IFN-γ) is one kind of crucial inflammatory cytokines, and its expression level is closely associated with various disease progressions. This work addresses the development of a sensitive and specific electrochemical assay for detection of IFN-γ by combing the recognition unit of aptamer with the signal reporter of target-induced silver nanoclusters (AgNCs). For biosensor preparation, the gold nanoparticles (AuNPs) immobilized on the amine-terminated electrode surface provided electrochemical interfaces for the self-assembly of C-rich modified aptamers. Then, the aptamer recognized IFN-γ and the free aptamer hybridized with conjugated DNA sequences. After the nuclease-catalyzed cleavage of DNA duplex, in situ-generated AgNCs in the C-rich template was utilized as the electrochemical indicator for IFN-γ detection. The present method demonstrated a good performance for detection of IFN-γ with a low detection limit of 1.7 pg mL-1. This aptasensor was verified to be applied for the evaluation of IFN-γ secreted by cell.