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1.
Mikrochim Acta ; 191(7): 367, 2024 Jun 04.
Article in English | MEDLINE | ID: mdl-38832980

ABSTRACT

An electrochemical aptasensor was used for the fast and sensitive detection of zearalenone (ZEN) based on the combination of Co3O4/MoS2/Au nanocomposites and the hybrid chain reaction (HCR). The glassy carbon electrode was coated with Co3O4/MoS2/Au nanomaterials to immobilize the ZEN-cDNA that had been bound with ZEN-Apt by the principle of base complementary pairing. In the absence of ZEN, the HCR could not be triggered because the ZEN-cDNA could not be exposed. After ZEN was added to the surface of the electrode, a complex structure was produced on the modified electrode by the combination of ZEN and ZEN-Apt. Therefore, the ZEN-cDNA can raise the HCR to produce the long-strand dsDNA structure. Due to the formation of dsDNA, the methylene blue (MB) could be inserted into the superstructure of branched DNA and the peak currents of the MB redox signal dramatically increased. So the concentration of ZEN could be detected by the change of signal intensity. Under optimized conditions, the developed electrochemical biosensing strategy showed an outstanding linear detection range of 1.0×10-10 mol/L to 1.0×10-6 mol/L, a low detection limit (LOD) of 8.5×10-11 mol/L with desirable selectivity and stability. Therefore, the fabricated platform possessed a great application potential in fields of food safety, medical detection, and drug analysis.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Cobalt , Disulfides , Electrochemical Techniques , Gold , Limit of Detection , Molybdenum , Nanocomposites , Oxides , Zearalenone , Zearalenone/analysis , Zearalenone/chemistry , Gold/chemistry , Nanocomposites/chemistry , Electrochemical Techniques/methods , Aptamers, Nucleotide/chemistry , Molybdenum/chemistry , Biosensing Techniques/methods , Cobalt/chemistry , Disulfides/chemistry , Oxides/chemistry , Electrodes , DNA/chemistry , Metal Nanoparticles/chemistry
2.
Anal Chim Acta ; 1306: 342585, 2024 Jun 01.
Article in English | MEDLINE | ID: mdl-38692786

ABSTRACT

Herein, we developed a convenient and versatile dual-mode electrochemiluminescence (ECL) and photoelectrochemistry (PEC) sensing radar for the detection of Prostate-specific antigen (PSA), which has important implications for detection of low-abundance disease-associated proteins. Cerium-based metal-organic framework (Ce-MOFs) were firstly modified on the electrode, showing well ECL and PEC property. In particular, a unique multifunctional Au@CdS quantum dots (QDs) probe loaded numerous QDs and antibody was fabricated, not only displaying strong ECL and PEC signals, but also having specific recognition to PSA. After the signal probe was linked to the electrode by immune reaction, much amplified signals of ECL and PEC were generated for double-mode detection of PSA. Therefore, this work proposed a multifunctional Au@CdS QDs signal probe with excellent ECL and PEC performance, and developed an ultrasensitive photoelectric biosensing platform for dual-mode detection, which provides an effective method for health monitoring of cancer patients.


Subject(s)
Cadmium Compounds , Electrochemical Techniques , Metal-Organic Frameworks , Prostate-Specific Antigen , Quantum Dots , Sulfides , Quantum Dots/chemistry , Cadmium Compounds/chemistry , Sulfides/chemistry , Humans , Prostate-Specific Antigen/analysis , Prostate-Specific Antigen/blood , Metal-Organic Frameworks/chemistry , Gold/chemistry , Cerium/chemistry , Biosensing Techniques , Photochemical Processes , Limit of Detection , Electrodes , Luminescent Measurements
3.
Anal Chim Acta ; 1306: 342617, 2024 Jun 01.
Article in English | MEDLINE | ID: mdl-38692789

ABSTRACT

BACKGROUND: Alpha-fetoprotein (AFP) is a fetal protein that can indicate congenital anomalies such as Down syndrome and spinal canal blockage when detected at abnormal levels in pregnant women. Current AFP detection methods rely on invasive blood or serum samples, which require sophisticated equipment. From the many solutions proposed, colorimetric paper-based assays excel in point-of-care settings. The concept of paper-based ELISA (p-ELISA) enhances traditional methods, aligning with the ASSURED criteria for diagnostics in resource-limited regions. Despite success in microfluidic paper-based assay devices, laser printing remains underexplored for p-ELISA. Additionally, modifying the paper surface provides an additional layer of sensitivity enhancement. RESULTS: In this study, we developed a novel laser-printed paper-based ELISA (LP-pELISA) for rapid, sensitive, and noninvasive detection of AFP in saliva samples. The LP-pELISA platform was fabricated by printing hydrophobic barriers on filter paper using a laser printer, followed by depositing hydroxyapatite (HAp) as an immobilization material for the antibodies. The colorimetric detection was achieved using AuNPs functionalized with anti-AFP antibodies and silver nitrate enhancement. The LP-pELISA exhibited a linear response for AFP detection in both buffer and saliva samples over a range of 1.0-800 ng mL-1, with a limit of detection (LOD) reaching 1.0 ng mL-1. The assay also demonstrated good selectivity, repeatability, reproducibility, and stability. The LP-pELISA was further validated by testing spiked human saliva samples, showing its potential for point-of-care diagnosis of congenital disabilities. SIGNIFICANCE: The LP-pELISA is a noninvasive platform showcasing simplicity, cost-effectiveness, and user-friendliness, utilizing laser printing, hydroxyapatite modification, and saliva samples to efficiently detect AFP. Beyond its application for AFP, this method's versatility extends to other biomarkers, positioning it as a catalyst for the evolution of paper-based biosensors. The LP-pELISA holds promise as a transformative tool for point-of-care diagnostics, fostering advancements in healthcare with its innovative technology.


Subject(s)
Colorimetry , Durapatite , Enzyme-Linked Immunosorbent Assay , Lasers , Paper , Saliva , alpha-Fetoproteins , Humans , Saliva/chemistry , Durapatite/chemistry , alpha-Fetoproteins/analysis , Printing , Gold/chemistry , Limit of Detection , Antibodies, Immobilized/immunology , Antibodies, Immobilized/chemistry
4.
Anal Chim Acta ; 1306: 342613, 2024 Jun 01.
Article in English | MEDLINE | ID: mdl-38692794

ABSTRACT

Glucose detection is of significant importance in providing information to the human health management. However, conventional enzymatic glucose sensors suffer from a limited long-term stability due to the losing activity of the enzymes. In this work, the AuNi bimetallic aerogel with a well-defined nanowire network is synthesized and applied as the sensing nanomaterial in the non-enzymatic glucose detection. The three-dimensional (3D) hierarchical porous structure of the AuNi bimetallic aerogel ensures the high sensitivity of the sensor (40.34 µA mM-1 cm-2). Theoretical investigation unveiled the mechanism of the boosting electrocatalytic activity of the AuNi bimetallic aerogel toward glucose. A better adhesion between the sensing nanomaterial and the screen-printing electrodes (SPEs) is obtained after the introduction of Ni. On the basis of a wide linearity in the range of 0.1-5 mM, an excellent selectivity, an outstanding long-term stability (90 days) as well as the help of the signal processing circuit and an M5stack development board, the as-prepared glucose sensor successfully realizes remote monitoring of the glucose concentration. We speculate that this work is favorable to motivating the technological innovations of the non-enzymatic glucose sensors and intelligent sensing devices.


Subject(s)
Biosensing Techniques , Electrochemical Techniques , Gels , Glucose , Gold , Nickel , Biosensing Techniques/methods , Nickel/chemistry , Gels/chemistry , Gold/chemistry , Glucose/analysis , Electrodes , Nanowires/chemistry , Humans , Limit of Detection
5.
Artif Cells Nanomed Biotechnol ; 52(1): 270-277, 2024 Dec.
Article in English | MEDLINE | ID: mdl-38696132

ABSTRACT

Spherical gold/polyacrylic acid (Au/PAA) polymer-inorganic Janus nanoparticles (JNPs) with simultaneous therapeutic and targeting functions were fabricated. The obtained Au/PAA JNPs were further selectively functionalized with folic acid (FA) and thiol PEG amine (SH-PEG-NH2) on Au sides to provide superior biocompatibility and active targeting, while the other PAA sides were loaded with 5-aminolevulinic acid (5-ALA) to serve as a photosensitizer (PS) for photodynamic therapeutic (PDT) effects on MCF-7 cancer cells. The PS loading of 5-ALA was found to be 83% with an average hydrodynamic size and z-potential of 146 ± 0.8 nm and -6.40 mV respectively for FA-Au/PAA-ALA JNPs. The in vitro PDT study of the JNPs on MCF-7 breast cancer cells under 636 nm laser irradiation indicated the cell viability of 24.7% ± 0.5 for FA-Au/PAA-ALA JNPs at the IC50 value of 0.125 mM. In this regard, the actively targeted FA-Au/PAA-ALA JNPs treatment holds great potential for tumour therapy with high cancer cell-killing efficacy.


Subject(s)
Aminolevulinic Acid , Breast Neoplasms , Gold , Photochemotherapy , Photosensitizing Agents , Humans , MCF-7 Cells , Breast Neoplasms/drug therapy , Breast Neoplasms/pathology , Breast Neoplasms/metabolism , Aminolevulinic Acid/chemistry , Aminolevulinic Acid/pharmacology , Gold/chemistry , Gold/pharmacology , Photosensitizing Agents/chemistry , Photosensitizing Agents/pharmacology , Nanoparticles/chemistry , Acrylic Resins/chemistry , Female , Folic Acid/chemistry , Cell Survival/drug effects
6.
Methods Cell Biol ; 187: 57-72, 2024.
Article in English | MEDLINE | ID: mdl-38705630

ABSTRACT

Correlative light and electron microscopy (CLEM) can provide valuable information about a biological sample by giving information on the specific localization of a molecule of interest within an ultrastructural context. In this work, we describe a simple CLEM method to obtain high-resolution images of neurotransmitter receptor distribution in synapses by electron microscopy (EM). We use hippocampal organotypic slices from a previously reported mouse model expressing a modified AMPA receptor (AMPAR) subunit that binds biotin at the surface (Getz et al., 2022). This tag can be recognized by StreptAvidin-Fluoronanogold™ conjugates (SA-FNG), which reach receptors at synapses (synaptic cleft is 50-100nm thick). By using pre-embedding labeling, we found that SA-FNG reliably bind synaptic receptors and penetrate around 10-15µm in depth in live tissue. However, the silver enhancement was only reaching the surface of the slices. We show that permeabilization with triton is highly effective at increasing the in depth-gold amplification and that the membrane integrity is well preserved. Finally, we also apply high-resolution electron tomography, thus providing important information about the 3D organization of surface AMPA receptors in synapses at the nanoscale.


Subject(s)
Hippocampus , Receptors, AMPA , Synapses , Animals , Mice , Hippocampus/metabolism , Hippocampus/cytology , Receptors, AMPA/metabolism , Synapses/metabolism , Synapses/ultrastructure , Membrane Proteins/metabolism , Gold/chemistry , Microscopy, Electron/methods , Fluorescent Dyes/chemistry , Fluorescent Dyes/metabolism
7.
Mikrochim Acta ; 191(6): 305, 2024 05 07.
Article in English | MEDLINE | ID: mdl-38713444

ABSTRACT

A multifunctional surface-enhanced Raman scattering (SERS) platform integrating sensitive detection and drug resistance analysis was developed for Gram-positive bacteria. The substrate was based on self-assembled Ti3C2Tx@Au NPs films and capture molecule phytic acid (IP6) to achieve specific capture of Gram-positive bacteria and different bacteria were analyzed by fingerprint signal. It had advantages of good stability and homogeneity (RSD = 8.88%). The detection limit (LOD) was 102 CFU/mL for Staphylococcus aureus and 103 CFU/mL for MRSA, respectively. A sandwich structure was formed on the capture substrate by signal labels prepared by antibiotics (penicillin G and vancomycin) and non-interference SERS probe molecules (4-mercaptobenzonitrile (2223 cm-1) and 2-amino-4-cyanopyridine (2240 cm-1)) to improve sensitivity. The LOD of Au NPs@4-MBN@PG to S. aureus and Au NPs@AMCP@Van to MRSA and S. aureus were all improved to 10 CFU/mL, with a wide dynamic linear range from 108 to 10 CFU/mL (R2 ≥ 0.992). The SERS platform can analyze the drug resistance of drug-resistant bacteria. Au NPs@4-MBN@PG was added to the substrate and captured MRSA to compare the SERS spectra of 4-MBN. The intensity inhomogeneity of 4-MBN at the same concentrations of MRSA and the nonlinearity at the different concentrations of MRSA revealed that MRSA was resistant to PG. Finally, the SERS platform achieved the determination of MRSA in blood. Therefore, this SERS platform has great significance for the determination and analysis of Gram-positive bacteria.


Subject(s)
Anti-Bacterial Agents , Gold , Limit of Detection , Metal Nanoparticles , Spectrum Analysis, Raman , Staphylococcus aureus , Titanium , Spectrum Analysis, Raman/methods , Gold/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Titanium/chemistry , Metal Nanoparticles/chemistry , Staphylococcus aureus/drug effects , Staphylococcus aureus/isolation & purification , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Vancomycin/pharmacology , Vancomycin/chemistry , Drug Resistance, Bacterial , Microbial Sensitivity Tests , Penicillin G/pharmacology , Penicillin G/chemistry , Gram-Positive Bacteria/drug effects , Gram-Positive Bacteria/isolation & purification
8.
Mikrochim Acta ; 191(5): 293, 2024 05 01.
Article in English | MEDLINE | ID: mdl-38691169

ABSTRACT

To address the need for facile, rapid detection of pathogens in water supplies, a fluorescent sensing array platform based on antibiotic-stabilized metal nanoclusters was developed for the multiplex detection of pathogens. Using five common antibiotics, eight different nanoclusters (NCs) were synthesized including ampicillin stabilized copper NCs, cefepime stabilized gold and copper NCs, kanamycin stabilized gold and copper NCs, lysozyme stabilized gold NCs, and vancomycin stabilized gold/silver and copper NCs. Based on the different interaction of each NC with the bacteria strains, unique patterns were generated. Various machine learning algorithms were employed for pattern discernment, among which the artificial neural networks proved to have the highest performance, with an accuracy of 100%. The developed prediction model performed well on an independent test dataset and on real samples gathered from drinking water, tap water and the Anzali Lagoon water, with prediction accuracy of 96.88% and 95.14%, respectively. This work demonstrates how generic antibiotics can be implemented for NC synthesis and used as recognition elements for pathogen detection. Furthermore, it displays how merging machine learning techniques can elevate sensitivity of analytical devices.


Subject(s)
Anti-Bacterial Agents , Copper , Gold , Metal Nanoparticles , Silver , Metal Nanoparticles/chemistry , Anti-Bacterial Agents/analysis , Anti-Bacterial Agents/chemistry , Gold/chemistry , Copper/chemistry , Silver/chemistry , Drinking Water/microbiology , Drinking Water/analysis , Neural Networks, Computer , Spectrometry, Fluorescence/methods , Machine Learning , Bacteria/isolation & purification , Fluorescent Dyes/chemistry , Vancomycin/chemistry , Water Microbiology , Kanamycin/analysis
9.
Mikrochim Acta ; 191(5): 294, 2024 05 02.
Article in English | MEDLINE | ID: mdl-38698253

ABSTRACT

Early transition metal carbides (MXene) hybridized by precious metals open a door for innovative electrochemical biosensing device design. Herein, we present a facile one-pot synthesis of gold nanoparticles (AuNPs)-doped two-dimensional (2D) titanium carbide MXene nanoflakes (Ti3C2Tx/Au). Ti3C2Tx MXene exhibits high electrical conductivity and yields synergistic signal amplification in conjunction with AuNPs leading to excellent electrochemical performance. Thus Ti3C2Tx/Au hybrid nanostructure can be used as an electrode platform for the electrochemical analysis of various targets. We used screen-printed electrodes modified with the Ti3C2Tx/Au electrode and functionalized with different biorecognition elements to detect and quantify an antibiotic, ampicillin (AMP), and a mycotoxin, fumonisin B1 (FB1). The ultralow limits of detection of 2.284 pM and 1.617 pg.mL-1, which we achieved respectively for AMP and FB1 are far lower than their corresponding maximum residue limits of 2.8 nM in milk and 2 to 4 mg kg-1 in corn products for human consumption set by the United States Food and Drug Administration. Additionally, the linear range of detection and quantification of AMP and FB1 were, respectively, 10 pM to 500 nM and 10 pg mL-1 to 1 µg mL-1. The unique structure and excellent electrochemical performance of Ti3C2Tx/Au nanocomposite suggest that it is highly suitable for anchoring biorecognition entities such as antibodies and oligonucleotides for monitoring various deleterious contaminants in agri-food products.


Subject(s)
Ampicillin , Electrochemical Techniques , Fumonisins , Gold , Limit of Detection , Metal Nanoparticles , Titanium , Fumonisins/analysis , Gold/chemistry , Ampicillin/analysis , Ampicillin/chemistry , Metal Nanoparticles/chemistry , Electrochemical Techniques/methods , Electrochemical Techniques/instrumentation , Titanium/chemistry , Biosensing Techniques/methods , Milk/chemistry , Anti-Bacterial Agents/analysis , Electrodes , Food Contamination/analysis , Animals
10.
Transl Vis Sci Technol ; 13(5): 5, 2024 May 01.
Article in English | MEDLINE | ID: mdl-38713474

ABSTRACT

Purpose: The blood-retinal barrier (BRB) restricts the delivery of intravenous therapeutics to the retina, necessitating innovative approaches for treating retinal disorders. This study sought to explore the potential of focused ultrasound (FUS) to non-invasively deliver intravenously administered gold nanoparticles (AuNPs) across the BRB. FUS-BRB modulation can offer a novel method for targeted retinal therapy. Methods: AuNPs of different sizes and shapes were characterized, and FUS parameters were optimized to permeate the BRB without causing retinal damage in a rodent model. The delivery of 70-kDa dextran and AuNPs to the retinal ganglion cell (RGC) layer was visualized using confocal and two-photon microscopy, respectively. Histological and statistical analyses were conducted to assess the effectiveness and safety of the procedure. Results: FUS-BRB modulation resulted in the delivery of dextran and AuNPs to the RGC and inner nuclear layer. Smaller AuNPs reached the retinal layers to a greater extent than larger ones. The delivery of dextran and AuNPs across the BRB with FUS was achieved without significant retinal damage. Conclusions: This investigation provides the first evidence, to our knowledge, of FUS-mediated AuNP delivery across the BRB, establishing a foundation for a targeted and non-invasive approach to retinal treatment. The results contribute to developing promising non-invasive therapeutic strategies in ophthalmology to treat retinal diseases. Translational Relevance: Modifying the BRB with ultrasound offers a targeted and non-invasive delivery strategy of intravenous therapeutics to the retina.


Subject(s)
Blood-Retinal Barrier , Gold , Metal Nanoparticles , Retinal Ganglion Cells , Animals , Gold/chemistry , Gold/administration & dosage , Retinal Ganglion Cells/cytology , Metal Nanoparticles/administration & dosage , Metal Nanoparticles/chemistry , Dextrans/administration & dosage , Dextrans/chemistry , Drug Delivery Systems/methods , Rats , Microscopy, Confocal/methods , Male
11.
Nanotechnology ; 35(33)2024 May 28.
Article in English | MEDLINE | ID: mdl-38701760

ABSTRACT

For the first time, this study shows the nanoarchitectonic process to obtain an acetogenin-enriched nanosystem (AuNPs-Ac) using an aqueous extract fromAnnona cherimolaMill (ACM) composed of gold nanoparticles embedded in an organic matrix that acts as stabilizing agent and presents anti-inflammatory activity and cytotoxical effect against HepG2 cell line, promoting apoptosis. The synthesis of AuNPs-Ac was confirmed by x-ray diffraction analysis, showing metallic gold as the only phase, and the scanning transmission microscope showed an organic cap covering the AuNPs-Ac. Fourier-transformed infrared suggests that the organic cap comprises a combination of different annonaceous acetogenins, alkaloids, and phenols by the presence of bands corresponding to aromatic rings and hydroxyl groups. High-Performance Liquid Chromatography has demonstrated the presence of annonacin, a potent acetogenin, in the extract of ACM. Anin vitroanti-inflammatory activity of the extract of ACM and the AuNPs-Ac was performed using the albumin denaturation method, showing a nonlinear response, which is better than sodium diclofenac salt in a wide range of concentrations that goes from 200 to 400µg ml-1with both samples. The viability assay was studied using trypan blue, treating IMR90 and HepG2 at different concentrations of AuNPs-Ac. The results defined a median lethal dose of 800µg ml-1against HepG2 through apoptosis according to the ratio of caspase-cleaved 9/alpha-tubulin evaluated. It was also demonstrated that the nanosystem presents a higher cytotoxic effect on the HepG2 cell line than in IMR90, suggesting a targeted mechanism. In addition, the nanosystem performs better than using only the extract of ACM in the anti-inflammatory or antiproliferative test, attributed to their higher surface area.


Subject(s)
Acetogenins , Anti-Inflammatory Agents , Apoptosis , Gold , Metal Nanoparticles , Plant Extracts , Humans , Acetogenins/pharmacology , Acetogenins/chemistry , Hep G2 Cells , Apoptosis/drug effects , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Metal Nanoparticles/chemistry , Gold/chemistry , Gold/pharmacology , Cell Survival/drug effects
12.
Biosens Bioelectron ; 258: 116356, 2024 Aug 15.
Article in English | MEDLINE | ID: mdl-38705073

ABSTRACT

In this work, the dual-ligand lanthanide metal-organic framework (MOF)-based electrochemiluminescence (ECL) sensor was constructed for the detection of miRNA-128 in glioblastoma (GBM) diagnosis. The luminescent Eu-MOF (EuBBN) was synthesized with terephthalic acid (BDC) and 2-amino terephthalic acid (BDC-NH2) as dual-ligand. Due to the antenna effect, EuBBN with conjugated-π structure exhibited strong luminescent signal and high quantum efficiency, which can be employed as ECL nanoprobe. Furthermore, the novel plasmonic CuS@Au heterostructure array has been prepared. The localized surface plasmon resonance coupling effect of the CuS@Au heterostructure array can amplify the ECL signal of EuBBN significantly. The EuBBN/CuS@Au heterostructure array-based sensing system has been prepared for the detection of miRNA-128 with a wide linear range from 1 fM to 1 nM and a detection limit of 0.24 fM. Finally, miRNA-128 in the clinic GBM tissue sample has been analysis for the distinguish of tumor grade successfully. The results demonstrated that the dual-ligand MOF/CuS@Au heterostructure array-based ECL sensor can provide important support for the development of GBM diagnosis.


Subject(s)
Biosensing Techniques , Europium , Glioblastoma , Gold , Metal-Organic Frameworks , MicroRNAs , MicroRNAs/analysis , Glioblastoma/diagnosis , Humans , Metal-Organic Frameworks/chemistry , Biosensing Techniques/methods , Gold/chemistry , Europium/chemistry , Limit of Detection , Luminescent Measurements/methods , Ligands , Electrochemical Techniques/methods , Brain Neoplasms/diagnosis , Phthalic Acids/chemistry , Metal Nanoparticles/chemistry , Copper/chemistry
13.
Anal Chem ; 96(21): 8390-8398, 2024 May 28.
Article in English | MEDLINE | ID: mdl-38716680

ABSTRACT

In this work, a microfluidic immunosensor chip was developed by incorporating microfluidic technology with electrochemiluminescence (ECL) for sensitive detection of human epidermal growth factor receptor-2 (HER2). The immunosensor chip can achieve robust reproducibility in mass production by integrating multiple detection units in a series. Notably, nanoscale materials can be better adapted to microfluidic systems, greatly enhancing the accuracy of the immunosensor chip. Ag@Au NCs closed by glutathione (GSH) were introduced in the ECL microfluidic immunosensor system with excellent and stable ECL performance. The synthesized CeO2-Au was applied as a coreaction promoter in the ECL signal amplification system, which made the result of HER2 detection more reliable. In addition, the designed microfluidic immunosensor chip integrated the biosensing system into a microchip, realizing rapid and accurate detection of HER2 by its high throughput and low usage. The developed short peptide ligand NARKFKG (NRK) achieved an effective connection between the antibody and nanocarrier for improving the detection efficiency of the sensor. The immunosensor chip had better storage stability and sensitivity than traditional detection methods, with a wide detection range from 10 fg·mL-1 to 100 ng·mL-1 and a low detection limit (LOD) of 3.29 fg·mL-1. In general, a microfluidic immunosensor platform was successfully constructed, providing a new idea for breast cancer (BC) clinical detection.


Subject(s)
Biosensing Techniques , Electrochemical Techniques , Electrodes , Gold , Luminescent Measurements , Metal Nanoparticles , Receptor, ErbB-2 , Silver , Humans , Receptor, ErbB-2/analysis , Receptor, ErbB-2/immunology , Metal Nanoparticles/chemistry , Electrochemical Techniques/methods , Silver/chemistry , Biosensing Techniques/methods , Gold/chemistry , Immunoassay/methods , Microfluidic Analytical Techniques/instrumentation , Limit of Detection , Cerium/chemistry
14.
Anal Chem ; 96(21): 8365-8372, 2024 May 28.
Article in English | MEDLINE | ID: mdl-38717986

ABSTRACT

Simultaneous sensitive and precise determination of multibiomarkers is of great significance for improving detection efficiency, reducing diagnosis and treatment expenses, and elevating survival rates. However, the development of simple and portable biosensors for simultaneous determination of multiplexed targets in biological fluids still faces challenges. Herein, a unique and versatile immobilization-free dual-target electrochemical biosensing platform, which combines distinguishable magnetic signal reporters with buoyancy-magnetism separation, was designed and constructed for simultaneous detection of carcinoembryonic (CEA) and α-fetoprotein (AFP) in intricate biological fluids. To construct such distinguishable magnetic signal reporters with signal transduction, amplification, and output, secondary antibodies of CEA and AFP were respectively functionalized on methylene blue (MB) and 6-(ferrocenyl)hexanethiol (FeC) modified Fe3O4@Au magnetic nanocomposites. Meanwhile, a multifunctional flotation probe with dual target recognition, capture, and isolation capability was prepared by conjugating primary antibodies (Ab1-CEA, Ab1-AFP) to hollow buoyant microspheres. The target antigens of CEA and AFP can trigger a flotation-mediated sandwich-type immunoreaction and capture a certain amount of the distinguishable magnetic signal reporter, which enables the conversion of the target CEA and AFP quantities to the signal of the potential-resolved MB and FeC. Thus, the MB and FeC currents of magnetically adsorbed distinguishable magnetic reporters can be used to determine the CEA and AFP targets simultaneously and precisely. Accordingly, the proposed strategy exhibited a delightful linear response for CEA and AFP in the range of 100 fg·mL-1-100 ng·mL-1 with detection limits of 33.34 and 17.02 fg·mL-1 (S/N = 3), respectively. Meanwhile, no significant nonspecific adsorption and cross-talk were observed. The biosensing platform has shown satisfactory performance in the determination of real clinical samples. More importantly, the proposed approach can be conveniently extended to universal detection just by simply substituting biorecognition events. Thus, this work opens up a new promising perspective for dual and even multiple targets and offers promising potential applications in clinical diagnosis.


Subject(s)
Biosensing Techniques , Carcinoembryonic Antigen , Electrochemical Techniques , alpha-Fetoproteins , alpha-Fetoproteins/analysis , alpha-Fetoproteins/immunology , Carcinoembryonic Antigen/analysis , Carcinoembryonic Antigen/immunology , Biosensing Techniques/methods , Humans , Immunoassay/methods , Gold/chemistry , Limit of Detection
15.
Biosens Bioelectron ; 258: 116340, 2024 Aug 15.
Article in English | MEDLINE | ID: mdl-38718633

ABSTRACT

The escalating global incidence of infectious diseases caused by pathogenic bacteria, especially in developing countries, emphasises the urgent need for rapid and portable pathogen detection devices. This study introduces a sensitive and specific electrochemical biosensing platform utilising cost-effective electrodes fabricated by inkjet-printing gold and silver nanoparticles on a plastic substrate. The biosensor exploits the CRISPR/Cas12a system for detecting a specific DNA sequence selected from the genome of the target pathogen. Upon detection, the trans-activity of Cas12a/gRNA is triggered, leading to the cleavage of rationally designed single-strand DNA reporters (linear and hairpin) labelled with methylene blue (ssDNA-MB) and bound to the electrode surface. In principle, this sensing mechanism can be adapted to any bacterium by choosing a proper guide RNA to target a specific sequence of its DNA. The biosensor's performance was assessed for two representative pathogens (a Gram-negative, Escherichia coli, and a Gram-positive, Staphylococcus aureus), and results obtained with inkjet-printed gold electrodes were compared with those obtained by commercial screen-printed gold electrodes. Our results show that the use of inkjet-printed nanostructured gold electrodes, which provide a large surface area, in combination with the use of hairpin reporters containing a poly-T loop can increase the sensitivity of the assay corresponding to a signal variation of 86%. DNA targets amplified from various clinically isolated bacteria, have been tested and demonstrate the potential of the proposed platform for point-of-need applications.


Subject(s)
Biosensing Techniques , CRISPR-Cas Systems , Escherichia coli , Gold , Metal Nanoparticles , Staphylococcus aureus , Biosensing Techniques/instrumentation , Gold/chemistry , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/genetics , Escherichia coli/isolation & purification , Escherichia coli/genetics , Metal Nanoparticles/chemistry , Silver/chemistry , DNA, Bacterial/analysis , DNA, Bacterial/genetics , Electrochemical Techniques/methods , Humans , Nanostructures/chemistry , DNA, Single-Stranded/chemistry , Electrodes , Printing , Bacterial Proteins/genetics , Endodeoxyribonucleases , CRISPR-Associated Proteins
16.
Anal Chem ; 96(21): 8682-8688, 2024 May 28.
Article in English | MEDLINE | ID: mdl-38757179

ABSTRACT

Programming ultrasensitive and stimuli-responsive DNAzyme-based probes holds great potential for on-demand biomarker detection. Here, an optically triggered DNAzyme platform was reported for on-demand activation-sensitive electrochemiluminescence (ECL) c-myc mRNA analysis. In this design, the sensing and recognition function of the split DNAzyme (SDz) probe was silent by engineering a blocking sequence containing a photocleavable linker (PC-linker) group at a defined site that could be indirectly cleaved by 302 nm ultraviolet (UV) light. When the SDz probes were assembled on the Au nanoparticles and potassium (K) element doped graphitic carbon nitride nanosheet (K-doped g-C3N4) covered electrode, UV light activation induces the configurational switching and consequently the formation of an active DNAzyme probe with the help of target c-myc mRNA, allowing the cleavage of the substrate strand by magnesium ions (Mg2+). Thus, the release of a ferrocene (Fc)-labeled DNAzyme 2 strand contributed to an extreme ECL signal recovery. In the meantime, the released target c-myc mRNA combined another inactive SDz motif to form active DNAzyme and repeat the cyclic cleavage reaction, resulting in the signal amplification. Furthermore, according to the responses toward two other designed nPC-SDz and m-SDz probes, we demonstrated that controlled UV light mediated photoactivation of the DNAzyme biosensor "on demand" effectively constrained the ECL signal to the mRNA of interest. Moreover, false positive signals could also be avoided due to such a photoactivation design with UV light. Therefore, this study provided a simple methodology that may be broadly applicable for investigating the mRNA-associated physiological events that were difficult to access using traditional DNAzyme probes.


Subject(s)
DNA, Catalytic , Electrochemical Techniques , Luminescent Measurements , RNA, Messenger , DNA, Catalytic/metabolism , DNA, Catalytic/chemistry , Electrochemical Techniques/methods , RNA, Messenger/analysis , Humans , Ultraviolet Rays , Biosensing Techniques/methods , Gold/chemistry , Metal Nanoparticles/chemistry , Metal Nanoparticles/radiation effects , Photochemical Processes , Proto-Oncogene Proteins c-myc/genetics , Proto-Oncogene Proteins c-myc/metabolism , Graphite/chemistry , Limit of Detection , Nitrogen Compounds
17.
Molecules ; 29(10)2024 May 19.
Article in English | MEDLINE | ID: mdl-38792255

ABSTRACT

The rapid advancements in nanotechnology in the field of nanomedicine have the potential to significantly enhance therapeutic strategies for cancer treatment. There is considerable promise for enhancing the efficacy of cancer therapy through the manufacture of innovative nanocomposite materials. Metallic nanoparticles have been found to enhance the release of anticancer medications that are loaded onto them, resulting in a sustained release, hence reducing the dosage required for drug administration and preventing their buildup in healthy cells. The combination of nanotechnology with biocompatible materials offers new prospects for the development of advanced therapies that exhibit enhanced selectivity, reduced adverse effects, and improved patient outcomes. Chitosan (CS), a polysaccharide possessing distinct physicochemical properties, exhibits favorable attributes for controlled drug delivery due to its biocompatibility and biodegradability. Chitosan nanocomposites exhibit heightened stability, improved biocompatibility, and prolonged release characteristics for anticancer medicines. The incorporation of gold (Au) nanoparticles into the chitosan nanocomposite results in the manifestation of photothermal characteristics, whereas the inclusion of silver (Ag) nanoparticles boosts the antibacterial capabilities of the synthesized nanocomposite. The objective of this review is to investigate the recent progress in the utilization of Ag and Au nanoparticles, or a combination thereof, within a chitosan matrix or its modified derivatives for the purpose of anticancer drug delivery. The research findings for the potential of a chitosan nanocomposite to deliver various anticancer drugs, such as doxorubicin, 5-Fluroacil, curcumin, paclitaxel, and 6-mercaptopurine, were investigated. Moreover, various modifications carried out on the chitosan matrix phase and the nanocomposite surfaces to enhance targeting selectivity, loading efficiency, and pH sensitivity were highlighted. In addition, challenges and perspectives that could motivate further research related to the applications of chitosan nanocomposites in cancer therapy were summarized.


Subject(s)
Antineoplastic Agents , Chitosan , Gold , Metal Nanoparticles , Nanocomposites , Silver , Chitosan/chemistry , Nanocomposites/chemistry , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Antineoplastic Agents/administration & dosage , Silver/chemistry , Humans , Metal Nanoparticles/chemistry , Gold/chemistry , Drug Delivery Systems , Drug Carriers/chemistry , Neoplasms/drug therapy , Animals
18.
Mikrochim Acta ; 191(6): 299, 2024 05 06.
Article in English | MEDLINE | ID: mdl-38709371

ABSTRACT

Gold nanoclusters are a smart platform for sensing potassium ions (K+). They have been synthesized using bovine serum albumin (BSA) and valinomycin (Val) to protect and cap the nanoclusters. The nanoclusters (Val-AuNCs) produced have a red emission at 616 nm under excitation with 470 nm. In the presence of K+, the valinomycin polar groups switch to the molecule's interior by complexing with K+, forming a bracelet structure, and being surrounded by the hydrophobic exterior conformation. This structure allows a proposed fluorometric method for detecting K+ by switching between the Val-AuNCs' hydrophilicity and hydrophobicity, which induces the aggregation of gold nanoclusters. As a result, significant quenching is seen in fluorescence after adding K+. The quenching in fluorescence in the presence of K+ is attributed to the aggregation mechanism. This sensing technique provides a highly precise and selective sensing method for K+ in the range 0.78 to 8 µM with LOD equal to 233 nM. The selectivity of Val-AuNCs toward K+ ions was investigated compared to other ions. Furthermore, the Val-AuNCs have novel possibilities as favorable sensor candidates for various imaging applications. Our detection technique was validated by determining K+ ions in postmortem vitreous humor samples, which yielded promising results.


Subject(s)
Fluorescent Dyes , Gold , Metal Nanoparticles , Potassium , Serum Albumin, Bovine , Valinomycin , Gold/chemistry , Valinomycin/chemistry , Potassium/analysis , Potassium/chemistry , Metal Nanoparticles/chemistry , Serum Albumin, Bovine/chemistry , Fluorescent Dyes/chemistry , Spectrometry, Fluorescence/methods , Limit of Detection , Animals , Hydrophobic and Hydrophilic Interactions , Cattle
19.
Mikrochim Acta ; 191(6): 298, 2024 05 06.
Article in English | MEDLINE | ID: mdl-38709403

ABSTRACT

As a real-time fluid biopsy method, the detection of circulating tumor cells (CTCs) provides important information for the early diagnosis, precise treatment, and prognosis of cancer. However, the low density of CTCs in the peripheral blood hampers their capture and detection with high sensitivity and selectivity using currently available methods. Hence, we designed a sandwich-type electrochemical aptasensor that utilizes holothurian-shaped AuPd nanoparticles (AuPd HSs), tetrahedral DNA nanostructures (TDNs), and CuPdPt nanowire networks (NWs) interwoven with a graphdiyne (GDY) sheet for ultrasensitive non-destructive detection of MCF-7 breast cancer cells. CuPdPt NW-GDY effectively enhanced the electron transfer rate and coupled with the loaded TDNs. The TDNs could capture MCF-7 cells with precision and firmness, and the resulting composite complex was combined with AuPd HSs to form a sandwich-type structure. This novel aptasensor showed a linear range between 10 and 106 cells mL-1 and an ultralow detection limit of 7 cells mL-1. The specificity, stability, and repeatability of the measurements were successfully verified. Moreover, we used benzonase nuclease to achieve non-destructive recovery of cells for further clinical studies. According to the results, our aptasensor was more sensitive measuring the number of CTCs than other approaches because of the employment of TDNs, CuPdPt NW-GDY, and AuPd HSs. We designed a reliable sensor system for the detection of CTCs in the peripheral blood, which could serve as a new approach for cancer diagnosis at an early stage.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , DNA , Electrochemical Techniques , Gold , Limit of Detection , Metal Nanoparticles , Neoplastic Cells, Circulating , Palladium , Neoplastic Cells, Circulating/pathology , Humans , MCF-7 Cells , Metal Nanoparticles/chemistry , Electrochemical Techniques/methods , Aptamers, Nucleotide/chemistry , Gold/chemistry , DNA/chemistry , Biosensing Techniques/methods , Palladium/chemistry
20.
Molecules ; 29(9)2024 Apr 27.
Article in English | MEDLINE | ID: mdl-38731508

ABSTRACT

This study delves into the physicochemical properties of inorganic hydroxyapatite (HAp) and hybrid hydroxyapatite-chitosan (HAp-CTS) granules, also gold-enriched, which can be used as aggregates in biomicroconcrete-type materials. The impact of granules' surface modifications with citric acid (CA) or polyethylene glycol (PEG) was assessed. Citric acid modification induced increased specific surface area and porosity in inorganic granules, contrasting with reduced parameters in hybrid granules. PEG modification resulted in a slight increase in specific surface area for inorganic granules and a substantial rise for hybrid granules with gold nanoparticles. Varied effects on open porosity were observed based on granule type. Microstructural analysis revealed increased roughness for inorganic granules post CA modification, while hybrid granules exhibited smoother surfaces. Novel biomicroconcretes, based on α-tricalcium phosphate (α-TCP) calcium phosphate cement and developed granules as aggregates within, were evaluated for compressive strength. Compressive strength assessments showcased significant enhancement with PEG modification, emphasizing its positive impact. Citric acid modification demonstrated variable effects, depending on granule composition. The incorporation of gold nanoparticles further enriched the multifaceted approach to enhancing calcium phosphate-based biomaterials for potential biomedical applications. This study demonstrates the pivotal role of surface modifications in tailoring the physicochemical properties of granules, paving the way for advanced biomicroconcretes with improved compressive strength for diverse biomedical applications.


Subject(s)
Citric Acid , Durapatite , Polyethylene Glycols , Citric Acid/chemistry , Durapatite/chemistry , Polyethylene Glycols/chemistry , Gold/chemistry , Biocompatible Materials/chemistry , Materials Testing , Chitosan/chemistry , Porosity , Metal Nanoparticles/chemistry , Chemical Phenomena , Compressive Strength , Surface Properties
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