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2.
Talanta ; 269: 125512, 2024 Mar 01.
Article in English | MEDLINE | ID: mdl-38091737

ABSTRACT

Measuring the levels of the biomarkers vanillylmandelic acid (VMA) and 5-Hydroxyindole-3-acetic acid (5-HIAA) is a valuable tool for clinical diagnosis not only of neuroblastoma or carcinoid syndrome, but also of essential hypertension, depression, migraine, and Tourette's syndrome. Herein, we explore using graphene quantum dots (GQDs) coated with molecularly imprinted polymer (MIP) as novel dual-imprinted sensors for selective and simultaneous determination of VMA and 5-HIAA in urine and plasma samples. The dual-MIP was successfully coated on the GQDs core via co-polymerization of (3-aminopropyl) triethoxysilane (APTES) and tetraethyl orthosilicate (TEOS), acting as functional and cross-linking monomers, respectively. In addition, we successfully created the dual imprinted VMA and 5-HIAA shell on the GQDs' core via a one-pot synthesis. We fabricated a facile and ready-to-use Origami three-dimensional electrochemical paper-based analytical device (Origami 3D-ePAD) for simultaneous determination of VMA and 5-HIAA using a GQDs@dual-MIP modified graphene electrode (GQDs@dual-MIP/SPGE). The Origami 3D-ePAD was designed to form a voltammetric cell on a three-layer foldable sheet with several advantages. For example, they were quickly assembled and enhanced the device's physical durability with the hydrophobic backup sheet. The developed dual imprinted Origami 3D-ePAD leads to substantially enhanced sensitivity and selectivity to electrochemical signal amplification generated from increasing the electrode-specific surface area, electrocatalytic activity, and the large numbers of dual imprinted sites for VMA and 5-HIAA detection. The synthetic recognition sites are highly selective for 5-HIAA and VMA molecules with an imprinting factor of 8.46 and 7.10, respectively. Quantitative analysis relying on square wave voltammetry reveals excellent linear dynamic ranges of around 0.001-25 µM, with detection limits of 0.023 nM for 5-HIAA and 0.047 nM for VMA (3Sb, n = 3). The Origami 3D-ePAD provides high accuracy and precision (i.e., recovery values of 5-HIAA ranged from 82.98 to 98.40 %, and VMA ranged from 83.28 to 104.39 %), and RSD less than 4.37 %) in urine and plasma samples without any evidence of interference. Hence, it is well suited as a facile and ready-to-use disposable device for point-of-care testing. It is straightforward, cost-effective, reproducible, and stable. Furthermore, it allows for rapid analysis (analysis time ∼20s) useful in medical diagnosis and other relevant fields.


Subject(s)
Carcinoid Tumor , Graphite , Molecular Imprinting , Quantum Dots , Humans , Quantum Dots/chemistry , Molecularly Imprinted Polymers , Graphite/chemistry , Vanilmandelic Acid , Biomarkers, Tumor , Limit of Detection , Hydroxyindoleacetic Acid , Acetates , Molecular Imprinting/methods , Electrochemical Techniques/methods
3.
Tech Mess ; 90(12): 761-785, 2023 Dec.
Article in English | MEDLINE | ID: mdl-38046181

ABSTRACT

This review provides an overview on bio- and chemosensors based on a thermal transducer platform that monitors the thermal interface resistance R th between a solid chip and the supernatant liquid. The R th parameter responds in a surprisingly strong way to molecular-scale changes at the solid-liquid interface, which can be measured thermometrically, using for instance thermocouples in combination with a controllable heat source. In 2012, the effect was first observed during on-chip denaturation experiments on complementary and mismatched DNA duplexes that differ in their melting temperature. Since then, the concept is addressed as heat-transfer method, in short HTM, and numerous applications of the basic sensing principle were identified. Functionalizing the chip with bioreceptors such as molecularly imprinted polymers makes it possible to detect neurotransmitters, inflammation markers, viruses, and environmental pollutants. In combination with aptamer-type receptors, it is also possible to detect proteins at low concentrations. Changing the receptors to surface-imprinted polymers has opened up new possibilities for quantitative bacterial detection and identification in complex matrices. In receptor-free variants, HTM was successfully used to characterize lipid vesicles and eukaryotic cells (yeast strains, cancer cell lines), the latter showing spontaneous detachment under influence of the temperature gradient inherent to HTM. We will also address modifications to the original HTM technique such as M-HTM, inverted HTM, thermal wave transport analysis TWTA, and the hot-wire principle. The article concludes with an assessment of the possibilities and current limitations of the method, together with a technological forecast.

4.
Article in English | MEDLINE | ID: mdl-37884758

ABSTRACT

Around 30% of the scientific papers published on imprinted polymers describe the recognition of proteins, nucleic acids, viruses, and cells. The straightforward synthesis from only one up to six functional monomers and the simple integration into a sensor are significant advantages as compared with enzymes or antibodies. Furthermore, they can be synthesized against toxic substances and structures of low immunogenicity and allow multi-analyte measurements via multi-template synthesis. The affinity is sufficiently high for protein biomarkers, DNA, viruses, and cells. However, the cross-reactivity of highly abundant proteins is still a challenge.

5.
ACS Appl Mater Interfaces ; 15(18): 22471-22484, 2023 May 10.
Article in English | MEDLINE | ID: mdl-37125734

ABSTRACT

The aeronautical industry demands facile lightweight and low-cost solutions to address climate crisis challenges. Graphene can be a valid candidate to tackle these functionalities, although its upscalability remains difficult to achieve. Consequently, graphene-related materials (GRM) are gathering massive attention as top-down graphite exfoliation processes at the industrial scale are feasible and often employed. In this work, environmentally friendly produced partially oxidized graphene nanosheets (POGNs) reduced by green solvents such as l-Ascorbic Acid to rGNs are proposed to deliver functional coatings based on a glass fiber composite or coated Al2024 T3 for strategic R&D questions in the aeronautical industry, i.e., low energy production, de-icing, and water uptake. In detail, energy efficiency in rGNs production is assessed via response-surface modeling of the powder conductivity, hence proposing an optimized reduction window. De-Icing functionality is verified by measuring the stable electrothermal property of an rGNs based composite over 24 h, and water uptake is elucidated by evaluating electrochemical and corrosion properties. Moreover, a mathematical model is proposed to depict the relation between the layers' sheet resistance and applied rGNs mass per area, which extends the system to other graphene-related materials, conductive two-dimensional materials, and various substrates. To conclude, the proposed system based on rGNs and epoxy paves the way for future multifunctional coatings, able to enhance the resistance of surfaces, such as airplane wings, in a flight harsh environment.

6.
Sensors (Basel) ; 23(7)2023 Mar 28.
Article in English | MEDLINE | ID: mdl-37050585

ABSTRACT

Staphylococcus epidermidis (S. epidermidis) belongs to methicillin-resistant bacteria strains that cause severe disease in humans. Herein, molecularly imprinted polymer (MIP) nanoparticles resulting from solid-phase synthesis on entire cells were employed as a sensing material to identify the species. MIP nanoparticles revealed spherical shapes with diameters of approximately 70 nm to 200 nm in scanning electron microscopy (SEM), which atomic force microscopy (AFM) confirmed. The interaction between nanoparticles and bacteria was assessed using height image analysis in AFM. Selective binding between MIP nanoparticles and S. epidermidis leads to uneven surfaces on bacteria. The surface roughness of S. epidermidis cells was increased to approximately 6.3 ± 1.2 nm after binding to MIP nanoparticles from around 1 nm in the case of native cells. This binding behavior is selective: when exposing Escherichia coli and Bacillus subtilis to the same MIP nanoparticle solutions, one cannot observe binding. Fluorescence microscopy confirms both sensitivity and selectivity. Hence, the developed MIP nanoparticles are a promising approach to identify (pathogenic) bacteria species.


Subject(s)
Molecular Imprinting , Nanoparticles , Humans , Polymers/chemistry , Molecular Imprinting/methods , Nanoparticles/chemistry , Molecularly Imprinted Polymers , Microscopy, Atomic Force
7.
Materials (Basel) ; 16(4)2023 Feb 20.
Article in English | MEDLINE | ID: mdl-36837379

ABSTRACT

An airplane is statistically struck by lightning every year. The need for lightweight aircraft to reduce the production of carbon dioxide has significantly reduced the presence of metals in favour of composites, resulting in lower lightning strike protection efficiency. In this perspective, we critically review the state of technologies in lightning strike protection solutions based on carbon materials, graphene, and MXenes. Furthermore, we comment on possible future research directions in the field.

8.
Talanta ; 254: 124199, 2023 Mar 01.
Article in English | MEDLINE | ID: mdl-36549138

ABSTRACT

Oxidized low-density lipoprotein (oxLDL) is the leading cause of atherosclerosis and cardiovascular diseases. Here, we created a simple colorimetric assay for sensitive and specific determination of oxLDL using a selective aptamer coupled with salt-induced gold nanoparticle (AuNP) aggregation. The aptamer was chosen by Systematic Evolution of Ligands by Exponential Enrichment to obtain a novel selective sequence towards oxLDL (as 5'-CCATCACGGGGCAGGCGGACAAGGGGTAAGGGCCACATCA-3'). Mixing a 5 µM aptamer solution with an aliquot of a sample containing oxLDL followed by adding AuNP solution (OD = 1) and 80 mmol L-1 NaCl achieved rapid results within 19 min: linear response to oxLDL from 0.002 to 0.5 µmol L-1 with high selectivity, a recovery accuracy of 100-111% at the 95% confidence interval, and within-run and between-run precision of 1-6% and 1-5% coefficient variations, respectively. Artificial serum diluted at least 1:8 with distilled water, analyzed by the aptamer-based colorimetric assay, showed excellent correlation with conventional thiobarbituric acid reactive substances (TBARS) (R2 = 0.9792) as a rapid colorimetric method without the need for sample preparation other than dilution.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Metal Nanoparticles , Gold , Colorimetry/methods , Biosensing Techniques/methods , Lipoproteins, LDL , Sodium Chloride
9.
Talanta ; 254: 124202, 2023 Mar 01.
Article in English | MEDLINE | ID: mdl-36549139

ABSTRACT

Herein, we present a novel Origami 3D-µPAD for colorimetric carbaryl detection using a super-efficient catalyst, namely mesoporous silica-platinum nanoparticles coated with a molecularly imprinted polymer (MSN-PtNPs@MIP). Morphological and structural characterization reveals that coating MIP on the MSN-PtNPs surface significantly increases the selective area, leading to larger numbers of imprinting sites for improved sensitivity and selectivity in determining carbaryl. The as-prepared MSN-PtNPs@MIP was used for catalytic oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by H2O2. Carbaryl selectively binds to the cavities embedded on the MSN-PtNPs surface and subsequently inhibits TMB oxidation leading the color to change to light blue. The change of reaction color from dark blue to light blue depends on the concentration of carbaryl within the 3D-µPAD detection zone. This design integrates the advantages of highly efficient sample delivery through micro channels (top layer) and efficient partition/separation paths (bottom layer) of the cellulose substrate to achieve both improved detection sensitivity and selectivity. Assay on the Origami 3D-µPAD can determine carbaryl by ImageJ detection, over a dynamic range of 0.002-20.00 mg kg-1, with a very low limit of detection at 1.5 ng g-1. The developed 3D-µPAD exhibit high accuracy when applied to detect carbaryl in fruits, with satisfactory recoveries from 90.1% to 104.0% and relative differences from the reference HPLC values less than 5.0%. Furthermore, the fabricated Origami 3D-µPAD provides reliable durability and good reproducibility (3.19% RSD for fifteen devices).


Subject(s)
Metal Nanoparticles , Molecular Imprinting , Carbaryl , Molecularly Imprinted Polymers , Silicon Dioxide/chemistry , Polymers/chemistry , Platinum , Metal Nanoparticles/chemistry , Hydrogen Peroxide , Microfluidics , Reproducibility of Results
10.
Anal Chem ; 94(48): 16692-16700, 2022 12 06.
Article in English | MEDLINE | ID: mdl-36409323

ABSTRACT

Nanoscale imprinting significantly increases the specific surface area and recognition capabilities of a molecularly imprinted polymer by improving accessibility to analytes, binding kinetics, and template removal. Herein, we present a novel synthetic route for a dual molecularly imprinted polymer (dual-MIP) of the carcinogen oxidative stress biomarkers 3-nitrotyrosine (3-NT) and 4-nitroquinolin-N-oxide (4-NQO) as coatings on graphene quantum-dot capped gold nanoparticles (GQDs-AuNPs). The dual-MIP was successfully coated on the GQDs-AuNPs core via a (3-mercaptopropyl) trimethoxysilane (MPTMS) linkage and copolymerization with the 3-aminopropyltriethoxysilane (APTMS) functional monomer. In addition, we fabricated a facile and compact three-dimensional electrochemical paper-based analytical device (3D-ePAD) for the simultaneous determination of the dual biomarkers using a GQDs-AuNPs@dual-MIP-modified graphene electrode (GQDs-AuNPs@dual-MIP/SPGE). The developed dual-MIP device provides greatly enhanced electrochemical signal amplification due to the improved electrode-specific surface area, electrocatalytic activity, and the inclusion of large numbers of dual-imprinted sites for 3-NT and 4-NQO detection. Quantitative analysis used square wave voltammetry, with an oxidation current appearing at -0.10 V for 4-NQO and +0.78 V for 3-NT. The dual-MIP sensor revealed excellent linear dynamic ranges of 0.01 to 500 µM for 3-NT and 0.005 to 250 µM for 4-NQO, with detection limits in nanomolar levels for both biomarkers. Furthermore, the dual-MIP sensor for the simultaneous determination of 3-NT and 4-NQO provides high accuracy and precision, with no evidence of interference from urine, serum, or whole blood samples.


Subject(s)
Graphite , Metal Nanoparticles , Molecular Imprinting , Gold , Molecularly Imprinted Polymers , Molecular Imprinting/methods , Electrochemical Techniques/methods , Carcinogens , Limit of Detection , Electrodes , Biomarkers , Oxidative Stress , Point-of-Care Testing
11.
Animals (Basel) ; 12(16)2022 Aug 14.
Article in English | MEDLINE | ID: mdl-36009664

ABSTRACT

Feline infectious peritonitis (FIP) is a worldwide fatal disease caused by a mutant feline coronavirus (FCoV). Simple and efficient molecular detection methods are needed. Here, sensitive, specific, rapid, and reliable colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed to detect the ORF1a/1b gene of FCoV from cats with suspected FIP using neutral red as an indicator. Novel LAMP primers were specifically designed based on the gene of interest. The isothermal assay could visually detect FCoV at 58 °C for 50 min. The RT-LAMP assay was highly specific and had no cross-reactivity with other related feline viruses. The detection limit of FCoV detection by RT-LAMP was 20 fg/µL. A blind clinical test (n = 81) of the developed RT-LAMP procedure was in good agreement with the conventional PCR method. In the light of its performance specificity, sensitivity, and easy visualization, this neutral-red-based RT-LAMP approach would be a fruitful alternative molecular diagnostic tool for veterinary inspection of FCoV when combined with nucleotide sequencing or specific PCR to affirm the highly virulent FIP-associated FCoV.

12.
Materials (Basel) ; 15(13)2022 Jul 01.
Article in English | MEDLINE | ID: mdl-35806768

ABSTRACT

To reduce the global emissions of CO2, the aviation industry largely relies on new light weight materials, which require multifunctional coatings. Graphene and its derivatives are particularly promising for combining light weight applications with functional coatings. Although they have proven to have outstanding properties, graphene and its precursor graphene oxide (GO) remain far from application at the industrial scale since a comprehensive protocol for mass production is still lacking. In this work, we develop and systematically describe a sustainable up-scaling process for the production of GO based on a three-step electrochemical exfoliation method. Surface characterization techniques (XRD, XPS and Raman) allow the understanding of the fast exfoliation rates obtained, and of high conductivities that are up to four orders of magnitude higher compared to GO produced via the commonly used modified Hummers method. Furthermore, we show that a newly developed mild thermal reduction at 250 °C is sufficient to increase conductivity by another order of magnitude, while limiting energy requirements. The proposed GO powder protocol suggests an up-scaling linear relation between the amount of educt surface and volume of electrolyte. This may support the mass production of GO-based coatings for the aviation industry, and address challenges such as low weight, fire, de-icing and lightning strike protection.

13.
Adv Sci (Weinh) ; 9(24): e2200459, 2022 08.
Article in English | MEDLINE | ID: mdl-35780480

ABSTRACT

Despite the importance of cell characterization and identification for diagnostic and therapeutic applications, developing fast and label-free methods without (bio)-chemical markers or surface-engineered receptors remains challenging. Here, we exploit the natural cellular response to mild thermal stimuli and propose a label- and receptor-free method for fast and facile cell characterization. Cell suspensions in a dedicated sensor are exposed to a temperature gradient, which stimulates synchronized and spontaneous cell-detachment with sharply defined time-patterns, a phenomenon unknown from literature. These patterns depend on metabolic activity (controlled through temperature, nutrients, and drugs) and provide a library of cell-type-specific indicators, allowing to distinguish several yeast strains as well as cancer cells. Under specific conditions, synchronized glycolytic-type oscillations are observed during detachment of mammalian and yeast-cell ensembles, providing additional cell-specific signatures. These findings suggest potential applications for cell viability analysis and for assessing the collective response of cancer cells to drugs.


Subject(s)
Eukaryotic Cells , Saccharomyces cerevisiae , Animals , Glycolysis , Mammals , Saccharomyces cerevisiae/metabolism
14.
J Mater Chem B ; 10(35): 6758-6767, 2022 09 15.
Article in English | MEDLINE | ID: mdl-35482413

ABSTRACT

Molecularly imprinted polymers (MIPs) are artificial recognition materials mimicking biological recognition entities such as antibodies. The general model of imprinting assumes that functional monomers interact with functional groups present on the target species which leads to cavities complementing the template in surface chemistry and shape thus ensuring recognition. However, to date there is little independent experimental evidence supporting that the surface chemistry in the imprints is tailored to analyte recognition and thus differs from the surface chemistry of the surrounding polymer. Herein, we investigate such chemical differences between imprints of Escherichia coli and Bacillus cereus in poly(styrene-co-DVB) and a commercial acrylate-based polymer by the means of confocal Raman microscopy and PLS-DA. The MIPs were generated using a stamping approach. Peak-force QNM measurements were conducted to rule out residues of bacterial cells in the imprints. While imprints of E. coli and B. cereus could be distinguished based on their Raman spectra in the acrylate-based polymer, differentiation in poly(styrene-co-DVB) was not significant. This could be a result of a higher potential of acrylate functional groups for interacting with lipopolysaccharides and peptidoglycans on bacteria surfaces compared to the phenyl groups of poly(styrene-co-DVB) and emphasizes the importance of the right choice of functional monomers for a specific target analyte.


Subject(s)
Molecular Imprinting , Acrylates , Escherichia coli , Microscopy, Scanning Probe , Molecularly Imprinted Polymers , Polymers/chemistry , Styrene
15.
Anal Chim Acta ; 1204: 339740, 2022 Apr 29.
Article in English | MEDLINE | ID: mdl-35397910

ABSTRACT

There is an urgent need for sensing strategies to screen perfluoroalkyl substances (PFAS) in aqueous matrices. These strategies must be applicable in large-scale monitoring plans to face the ubiquitous use of PFAS, their wide global spread, and their fast evolution towards short-chain, branched molecules. To this aim, the changes in fluorinated self-assembled monolayers (SAM) with different architectures (pinholes/defects-free and with randomized pinholes/defects) were studied upon exposure to both long and short-chain PFAS. The applicability of fluorinated SAM in PFAS sensing was evaluated. Changes in the SAM structures were characterised combining electrochemical impedance spectroscopy and voltammetric techniques. The experimental data interpretation was supported by molecular dynamics simulations to gain a more in-depth understanding of the interaction mechanisms involved. Pinhole/defect-free fluorinated SAM were found to be applicable to long-chain PFAS screening within switch-on sensing strategy, while a switch-off sensing strategy was reported for screening of both short/long-chain PFAS. These strategies confirmed the possibility to play on fluorophilic interactions when designing PFAS screening methods.


Subject(s)
Alkanesulfonic Acids , Fluorocarbons , Fluorocarbons/chemistry
16.
Soft Matter ; 18(11): 2245-2251, 2022 Mar 16.
Article in English | MEDLINE | ID: mdl-35234796

ABSTRACT

Investigations on lithographically formed cavities of surface-imprinted polymers (SIP) can help to gain deeper understanding on cell recognition with SIPs: it is known that surface topography and biomolecules transferred during surface imprinting contribute to cell adhesion. In this work, SIPs synthesized via two different imprinting techniques, namely stamp imprinting and polymerization of Pickering emulsions, were investigated and compared to each other, using atomic force microscopy (AFM) and Peak Force Quantitative Nano Mechanics (PF-QNM). We focused on SIPs based on poly(styrene-co-divinylbenzene) as model polymer and E. coli as model template for cell imprinting. Both imprinting approaches led to cavities that revealed nanostructures within the imprints. Stamp imprinting cavities feature low surface roughness and channel structures that resemble the negative pattern of the bacteria on the stamp and their filaments, while SIPs synthesized via polymerization of Pickering emulsions reveal globular nanostructures accumulating in the imprints. AFM phase imaging and adhesion mapping using PF-QNM show that these globular structures are remainders of the imprinted E. coli cells, most likely lipopolysaccarides, which is not observable in imprints resulting from stamp imprinting.


Subject(s)
Molecular Imprinting , Polymers , Escherichia coli , Microscopy, Atomic Force , Molecular Imprinting/methods , Polymerization , Polymers/chemistry
17.
ACS Appl Bio Mater ; 5(1): 160-171, 2022 01 17.
Article in English | MEDLINE | ID: mdl-35014817

ABSTRACT

Molecularly imprinted polymers (MIPs) are widely used as robust biomimetic recognition layers in sensing devices targeting a wide variety of analytes including microorganisms such as bacteria. Assessment of imprinting success and selectivity toward the target is of great importance in MIP quality control. We generated Escherichia coli-imprinted poly(styrene-co-DVB) as a model system for bacteria-imprinted polymers via surface imprinting using a glass stamp with covalently immobilized E. coli. Confocal Raman Microscopy was successfully employed to visualize bacteria, imprints, and polymer and to distinguish them from each other. The method has proven highly feasible for assessing if imprinting had been successful. In addition, we developed a method for selectivity investigation of bacteria MIPs based on combining Confocal Raman Microscopy and Partial Least Squares Discriminant Analysis (PLS-DA). The Raman spectra of E. coli and Bacillus cereus were acquired on E. coli-imprinted poly(styrene-co-DVB) and used to establish a PLS-DA model for differentiating between the bacteria species. Model validation demonstrated a correct classification of 95% of Raman spectra, indicating sufficient accuracy of the model for future use in MIP selectivity studies. Simultaneous differentiation of 3 bacteria species (E. coli, B. cereus, and Lactococcus lactis) on E. coli-imprinted poly(styrene-co-DVB) proved more difficult, which might be due to the limited depth resolution of the confocal Raman microscope resulting in the presence of interfering signals from the polymer substrate. It might be possible to overcome this obstacle by selective enhancement of the Raman signals originating from bacteria surfaces, such as tip enhanced Raman spectroscopy.


Subject(s)
Molecular Imprinting , Polymers , Escherichia coli , Molecular Imprinting/methods , Molecularly Imprinted Polymers , Polymers/chemistry , Spectrum Analysis, Raman/methods , Styrene
18.
Anal Chim Acta ; 1191: 339363, 2022 Jan 25.
Article in English | MEDLINE | ID: mdl-35033235

ABSTRACT

We present a novel dual-imprinted electrochemical paper-based analytical device (Di-ePAD) to simultaneously determine 8-hydroxy-2'-deoxyguanosine (8-OHdG) and 3-nitrotyrosine (3-NT) and assess oxidative and nitrative biomarkers in urine and plasma samples. The Di-ePAD was designed with hydrophobic barrier layers formed on filter paper to provide three-dimensional circular reservoirs and assembled electrodes. The molecularly imprinted polymer (MIP) was synthesized using a silica nanosphere decorated with silver nanoparticles (SiO2@AgNPs) as a core covered with dual-analyte imprinted sites on the polymer to recognize selectively and bind the target biomarkers. This strategy drives monodispersity and enhances the conductivity of the resulting MIP core-shell products. 3-NT-MIP and 8-OHdG-MIP were synthesized by successively coating the surface of SiO2@AgNPs with l-Cysteine via the thiol group, then terminating with MIP shells. The dual imprinted core-shell composites possess attractive properties for the target biomarkers' sensing, including catalytic activity, selectivity, and good conductivity. The Di-ePAD revealed excellent linear dynamic ranges of 0.01-500 µM for 3-NT and 0.05-500 µM for 8-OHdG, with detection limits of 0.0027 µM for 3-NT and 0.0138 µM for 8-OHdG. This newly developed method based on the synergistic effects of SiO2@AgNPs combined with promising properties of MIP offers outstanding selectivity, sensitivity, reproducibility, simplicity, and low cost for quantitative analysis of 3-NT and 8-OHdG. The proposed Di-ePAD showed good accuracy and precision when applied to actual samples, including urine and serum samples validated by a conventional HPLC method.


Subject(s)
Metal Nanoparticles , Molecular Imprinting , Biomarkers , Electrochemical Techniques , Electrodes , Limit of Detection , Oxidative Stress , Reproducibility of Results , Silicon Dioxide , Silver
19.
Anal Bioanal Chem ; 414(1): 731-741, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34950982

ABSTRACT

Solid-phase synthesis is an elegant way to create molecularly imprinted polymer nanoparticles (nano-MIPs) comprising a single binding site, i.e. mimics of antibodies. When using human serum albumin (HSA) as the template, one achieves nano-MIPs with 53 ± 19 nm diameter, while non-imprinted polymer nanoparticles (nano-NIPs) reach 191 ± 96 nm. Fluorescence assays lead to Stern-Volmer plots revealing selective binding to HSA with selectivity factors of 1.2 compared to bovine serum albumin (BSA), 1.9 for lysozyme, and 4.1 for pepsin. Direct quartz crystal microbalance (QCM) assays confirm these results: nano-MIPs bind to HSA immobilized on QCM surfaces. This opens the way for competitive QCM-based assays for HSA: adding HSA to nanoparticle solutions indeed reduces binding to the QCM surfaces in a concentration-dependent manner. They achieve a limit of detection (LoD) of 80 nM and a limit of quantification (LoQ) of 244 nM. Furthermore, the assay shows recovery rates around 100% for HSA even in the presence of competing analytes.


Subject(s)
Molecular Imprinting , Quartz Crystal Microbalance Techniques , Humans , Limit of Detection , Polymers/chemistry , Serum Albumin, Human
20.
Biosensors (Basel) ; 11(9)2021 Aug 31.
Article in English | MEDLINE | ID: mdl-34562899

ABSTRACT

Functionalized DNA sequences are promising sensing elements to combine with transducers for bio-sensing specific target microbes. As an application example, this paper demonstrates in situ detection of loop-mediated isothermal amplification products by hybridizing them with thiolated-ssDNA covalently anchored on the electrodes of a quartz crystal microbalance (QCM). Such hybridization leads to a frequency signal, which is suitable for monitoring real-time LAMP amplification based on mass-sensing: it detects interactions between the complementary nucleobases of LAMP products in solution and the thiolated-ssDNA probe sequence on the gold surface. Target DNA LAMP products cause irreversible frequency shifts on the QCM surfaces during hybridization in the kHz range, which result from both changes in mass and charge on the electrode surface. In order to confirm the LAMP assay working in the QCM sensing system at elevated temperature, the sky blue of positive LAMP products solution was achieved by using the Hydroxy Naphthol Blue (HNB) and agarose gel electrophoresis. Since on-QCM sensing of DNA hybridization leads to irreversible sensor responses, this work shows characterization by X-ray photoelectron spectroscopy (XPS) core spectra of S2p, N1s, Mg1s, P2p and C1s. XPS results confirmed that indeed both DNA and by-products of LAMP attached to the surface. Listeria monocytogenes DNA served to study in-situ detection of amplified LAMP products on DNA-functionalized surfaces.


Subject(s)
Listeria monocytogenes , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques , Listeria monocytogenes/genetics , Quartz Crystal Microbalance Techniques
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