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1.
Biosensors (Basel) ; 14(3)2024 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-38534241

RESUMO

Two types of low-cost reagentless electrochemical glucose biosensors based on graphite rod (GR) electrodes were developed. The electrodes modified with electrochemically synthesized platinum nanostructures (PtNS), 1,10-phenanthroline-5,6-dione (PD), glucose oxidase (GOx) without and with a polypyrrole (Ppy) layer-(i) GR/PtNS/PD/GOx and (ii) GR/PtNS/PD/GOx/Ppy, respectively, were prepared and tested. Glucose biosensors based on GR/PtNS/PD/GOx and GR/PtNS/PD/GOx/Ppy electrodes were characterized by the sensitivity of 10.1 and 5.31 µA/(mM cm2), linear range (LR) up to 16.5 and 39.0 mM, limit of detection (LOD) of 0.198 and 0.561 mM, good reproducibility, and storage stability. The developed glucose biosensors based on GR/PtNS/PD/GOx/Ppy electrodes showed exceptional resistance to interfering compounds and proved to be highly efficient for the determination of glucose levels in blood serum.


Assuntos
Técnicas Biossensoriais , Nanoestruturas , Glucose/química , Polímeros/química , Pirróis/química , Platina , Reprodutibilidade dos Testes , Eletrodos , Glucose Oxidase/química , Enzimas Imobilizadas/química
2.
Biosens Bioelectron ; 251: 116043, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38368643

RESUMO

This article aims to present a comparative study of three polypyrrole-based molecularly imprinted polymer (MIP) systems for the detection of the recombinant severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid protein (rN). The rN is known for its relatively low propensity to mutate compared to other SARS-CoV-2 antigens. The aforementioned systems include screen-printed carbon electrodes (SPCE) modified with gold nanostructures (MIP1), platinum nanostructures (MIP2), and the unmodified SPCE (MIP3), which was used for control. Pulsed amperometric detection (PAD) was employed as the detection technique, offering the advantage of label-free detection without the need for an additional redox probe. Calibration curves were constructed using the obtained data to evaluate the response of each system. Non-imprinted systems were also tested in parallel to evaluate the contribution of non-specific binding and assess the affinity sensor's efficiency. The analysis of calibration curves revealed that the AuNS-based MIP1 system exhibited the lowest contribution of non-specific binding and displayed a better fit with the chosen fitting model compared to the other systems. Further analysis of this system included determining the limit of detection (LOD) (51.2 ± 2.8 pg/mL), the limit of quantification (LOQ) (153.9 ± 8.3 pg/mL), and a specificity test using a recombinant receptor-binding domain of SARS-CoV-2 spike protein as a control. Based on the results, the AuNS-based MIP1 system demonstrated high specificity and sensitivity for the label-free detection of SARS-CoV-2 nucleocapsid protein. The utilization of PAD without the need for additional redox probes makes this sensing system convenient and valuable for rapid and accurate virus detection.


Assuntos
Técnicas Biossensoriais , COVID-19 , Glicoproteína da Espícula de Coronavírus , Humanos , SARS-CoV-2 , COVID-19/diagnóstico , Polímeros/química , Pirróis , Proteínas do Nucleocapsídeo/análise
3.
Biosensors (Basel) ; 13(7)2023 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-37504125

RESUMO

Reagentless electrochemical glucose biosensors were developed and investigated. A graphite rod (GR) electrode modified with electrochemically synthesized dendritic gold nanostructures (DGNs) and redox mediators (Med) such as ferrocenecarboxylic acid (FCA), 1,10-phenathroline-5,6-dione (PD), N,N,N',N'-tetramethylbenzidine (TMB) or tetrathiafulvalene (TTF) in combination with glucose oxidase (GOx) (GR/DGNs/FCA/GOx, GR/DGNs/PD/GOx, GR/DGNs/TMB/GOx, or GR/DGNs/TTF/GOx) were developed and electrochemically investigated. A biosensor based on threefold-layer-by-layer-deposited PD and GOx (GR/DGNs/(PD/GOx)3) was found to be the most suitable for the determination of glucose. To improve the performance of the developed biosensor, the surface of the GR/DGNs/(PD/GOx)3 electrode was modified with polypyrrole (Ppy) for 5 h. A glucose biosensor based on a GR/DGNs/(PD/GOx)3/Ppy(5 h) electrode was characterized using a wide linear dynamic range of up to 39.0 mmol L-1 of glucose, sensitivity of 3.03 µA mM-1 cm-2, limit of detection of 0.683 mmol L-1, and repeatability of 9.03% for a 29.4 mmol L-1 glucose concentration. The Ppy-based glucose biosensor was characterized by a good storage stability (τ1/2 = 9.0 days). Additionally, the performance of the developed biosensor in blood serum was investigated.


Assuntos
Técnicas Biossensoriais , Nanoestruturas , Glucose/química , Polímeros/química , Ouro/química , Enzimas Imobilizadas/química , Pirróis/química , Eletrodos , Glucose Oxidase/química
4.
Polymers (Basel) ; 15(11)2023 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-37299369

RESUMO

Bone marrow mesenchymal stem cells (BMMSCs) possess a strong ability to differentiate into the chondrogenic lineage, which is important for cartilage regeneration. External stimuli, such as electrical stimulation (ES), are frequently studied for chondrogenic differentiation of BMMSCs; however, the application of conductive polymers such as polypyrrole (Ppy), has never been used for stimulating BMMSCs chondrogenesis in vitro before. Thus, the aim of this study was to evaluate the chondrogenic potential of human BMMSCs after stimulation with Ppy nanoparticles (Ppy NPs) and compare them to cartilage-derived chondrocytes. In this study, we tested Ppy NPs without and with 13 nm gold NPs (Ppy/Au) for BMMSCs and chondrocyte proliferation, viability, and chondrogenic differentiation for 21 days, without the use of ES. The results demonstrated significantly higher amounts of cartilage oligomeric matrix protein (COMP) in BMMSCs stimulated with Ppy and Ppy/Au NPs, as compared to the control. The expression of chondrogenic genes (SOX9, ACAN, COL2A1) in BMMSCs and chondrocytes were upregulated by Ppy and Ppy/Au NPs, as compared to controls. Histological staining with safranin-O indicated higher extracellular matrix production in Ppy and Ppy/Au NPs stimulated samples, as compared to controls. In conclusion, Ppy and Ppy/Au NPs stimulate BMMSC chondrogenic differentiation; however, BMMSCs were more responsive to Ppy, while chondrocytes possessed a stronger chondrogenic response to Ppy/Au NPs.

5.
Polymers (Basel) ; 15(7)2023 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-37050211

RESUMO

Detecting bacteria-Listeria monocytogenes-is an essential healthcare and food industry issue. The objective of the current study was to apply platinum (Pt) and screen-printed carbon (SPCE) electrodes modified by molecularly imprinted polymer (MIP) in the design of an electrochemical sensor for the detection of Listeria monocytogenes. A sequence of potential pulses was used to perform the electrochemical deposition of the non-imprinted polypyrrole (NIP-Ppy) layer and Listeria monocytogenes-imprinted polypyrrole (MIP-Ppy) layer over SPCE and Pt electrodes. The bacteria were removed by incubating Ppy-modified electrodes in different extraction solutions (sulphuric acid, acetic acid, L-lysine, and trypsin) to determine the most efficient solution for extraction and to obtain a more sensitive and repeatable design of the sensor. The performance of MIP-Ppy- and NIP-Ppy-modified electrodes was evaluated by pulsed amperometric detection (PAD). According to the results of this research, it can be assumed that the most effective MIP-Ppy/SPCE sensor can be designed by removing bacteria with the proteolytic enzyme trypsin. The LOD and LOQ of the MIP-Ppy/SPCE were 70 CFU/mL and 210 CFU/mL, respectively, with a linear range from 300 to 6700 CFU/mL.

6.
Int J Mol Sci ; 24(6)2023 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-36982877

RESUMO

In this study, ZnO nanostructures with different types of morphologies and particle sizes were evaluated and applied for the development of an immunosensor. The first material was composed of spherical, polydisperse nanostructures with a particle size in the range of 10-160 nm. The second was made up of more compact rod-like spherical nanostructures with the diameter of these rods in the range of 50-400 nm, and approximately 98% of the particles were in the range of 20-70 nm. The last sample of ZnO was made up of rod-shaped particles with a diameter of 10-80 nm. These ZnO nanostructures were mixed with Nafion solution and drop-casted onto screen-printed carbon electrodes (SPCE), followed by a further immobilization of the prostate-specific antigen (PSA). The affinity interaction of PSA with monoclonal antibodies against PSA (anti-PSA) was evaluated using the differential pulse voltammetry technique. The limit of detection and limit of quantification of anti-PSA were determined as 1.35 nM and 4.08 nM for compact rod-shaped spherical ZnO nanostructures, and 2.36 nM and 7.15 nM for rod-shaped ZnO nanostructures, respectively.


Assuntos
Técnicas Biossensoriais , Óxido de Zinco , Humanos , Masculino , Anticorpos Monoclonais , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas , Eletrodos , Ouro/química , Imunoensaio/métodos , Limite de Detecção , Antígeno Prostático Específico/química , Óxido de Zinco/química , Nanopartículas Metálicas
7.
Biosensors (Basel) ; 12(8)2022 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-36004989

RESUMO

In this research, we assessed the applicability of electrochemical sensing techniques for detecting specific antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike proteins in the blood serum of patient samples following coronavirus disease 2019 (COVID-19). Herein, screen-printed carbon electrodes (SPCE) with electrodeposited gold nanostructures (AuNS) were modified with L-Cysteine for further covalent immobilization of recombinant SARS-CoV-2 spike proteins (rSpike). The affinity interactions of the rSpike protein with specific antibodies against this protein (anti-rSpike) were assessed using cyclic voltammetry (CV) and differential pulse voltammetry (DPV) methods. It was revealed that the SPCE electroactive surface area increased from 1.49 ± 0.02 cm2 to 1.82 ± 0.01 cm2 when AuNS were electrodeposited, and the value of the heterogeneous electron transfer rate constant (k0) changed from 6.30 × 10-5 to 14.56 × 10-5. The performance of the developed electrochemical immunosensor was evaluated by calculating the limit of detection and limit of quantification, giving values of 0.27 nM and 0.81 nM for CV and 0.14 nM and 0.42 nM for DPV. Furthermore, a specificity test was performed with a solution of antibodies against bovine serum albumin as the control aliquot, which was used to assess nonspecific binding, and this evaluation revealed that the developed rSpike-based sensor exhibits low nonspecific binding towards anti-rSpike antibodies.


Assuntos
Técnicas Biossensoriais , COVID-19 , Nanoestruturas , Anticorpos , Técnicas Biossensoriais/métodos , COVID-19/diagnóstico , Carbono/química , Técnicas Eletroquímicas/métodos , Eletrodos , Ouro/química , Humanos , Imunoensaio/métodos , Limite de Detecção , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus
8.
Biosensors (Basel) ; 12(8)2022 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-36005036

RESUMO

In this study, graphite rod (GR) electrodes were electrochemically modified by dendritic gold nanostructures (DGNs) followed by immobilization of glucose oxidase (GOx) in the presence of mediator phenazine methosulfate (PMS). Modified with polyaniline (PANI) or polypyrrole (Ppy), GOx/DGNs/GR electrodes were used in glucose biosensor design. Different electrochemical methods were applied for the registration of glucose concentration, and constant potential amperometry (CPA) was chosen as the best one. PANI and Ppy layers synthesized enzymatically on the GOx/DGNs/GR electrodes extended the linear glucose determination range, the width of which depended on the duration of PANI- and Ppy-layers formation. Enzymatically formed polypyrrole was determined as the most suitable polymer for the modification and formation of the glucose biosensor instead of polyaniline, because it was 1.35 times more sensitive and had a 2.57 times lower limit of detection (LOD). The developed glucose biosensor based on the Ppy/GOx/DGNs/GR electrode was characterized by appropriate sensitivity (59.4 µA mM-1 cm-2), low LOD (0.070 mmol L-1), wide linear glucose determination range (up to 19.9 mmol L-1), good repeatability (8.01%), and appropriate storage stability (33 days). The performance of the developed glucose biosensor was tested in biological samples and beverages.


Assuntos
Técnicas Biossensoriais , Grafite , Nanoestruturas , Técnicas Biossensoriais/métodos , Eletrodos , Enzimas Imobilizadas/química , Glucose , Glucose Oxidase/química , Ouro/química , Grafite/química , Polímeros/química , Pirróis/química
9.
Biosensors (Basel) ; 12(5)2022 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-35624652

RESUMO

SARS-CoV-2 vaccines provide strong protection against COVID-19. However, the emergence of SARS-CoV-2 variants has raised concerns about the efficacy of vaccines. In this study, we investigated the interactions of specific polyclonal human antibodies (pAb-SCoV2-S) produced after vaccination with the Vaxzevria vaccine with the spike proteins of three SARS-CoV-2 variants of concern: wild-type, B.1.1.7, and B.1.351. Highly sensitive, label-free, and real-time monitoring of these interactions was accomplished using the total internal reflection ellipsometry method. Thermodynamic parameters such as association and dissociation rate constants, the stable immune complex formation rate constant (kr), the equilibrium association and dissociation (KD) constants and steric factors (Ps) were calculated using a two-step irreversible binding mathematical model. The results obtained show that the KD values for the specific antibody interactions with all three types of spike protein are in the same nanomolar range. The KD values for B.1.1.7 and B.1.351 suggest that the antibody produced after vaccination can successfully protect the population from the alpha (B.1.1.7) and beta (B.1.351) SARS-CoV-2 mutations. The steric factors (Ps) obtained for all three types of spike proteins showed a 100-fold lower requirement for the formation of an immune complex when compared with nucleocapsid protein.


Assuntos
COVID-19 , Vacinas , Animais , Anticorpos Antivirais , Complexo Antígeno-Anticorpo , Vacinas contra COVID-19 , Humanos , Camundongos , Camundongos Endogâmicos BALB C , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/metabolismo
10.
Talanta ; 241: 123252, 2022 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-35121544

RESUMO

In this research the molecular imprinting technology was applied for the formation of glyphosate-sensitive layer. The glyphosate imprinted conducting polymer polypyrrole (MIPpy) was deposited on a gold chip/electrode and used as an electrochemical surface plasmon resonance (ESPR) sensor. The results described in this study disclose some restrictions and challenges, which arise during the development of glyphosate ESPR sensor based on the molecularly imprinted polymer development stage. It was demonstrated, that glyphosate could significantly affect the electrochemical deposition process of molecularly imprinted polymer on the electrode. The results of cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and surface plasmon resonance (SPR) have demonstrated that glyphosate molecules tend to interact with bare gold electrode and thus hinder the polypyrrole deposition. As a possible solution, the formation of a self-assembled monolayer (SAM) of 11-(1H-Pyrrol-1-yl)undecane-1-thiol (PUT) before electrochemical deposition of MIPpy and NIPpy was applied. Dissociation constant (KD) and free energy of Gibbs (ΔG0) values of glyphosate on MIPpy and Ppy without glyphosate imprints (NIPpy) were calculated. For the interaction of glyphosate with MIPpy the KD was determined as 38.18 ± 2.33⋅10-5 and ΔG0 as -19.51 ± 0.15 kJ/mol.


Assuntos
Impressão Molecular , Polímeros , Técnicas Eletroquímicas/métodos , Eletrodos , Glicina/análogos & derivados , Impressão Molecular/métodos , Polímeros/química , Pirróis/química , Ressonância de Plasmônio de Superfície , Glifosato
11.
Electrochim Acta ; 403: 139581, 2022 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-34898691

RESUMO

This study describes the application of a polypyrrole-based sensor for the determination of SARS-CoV-2-S spike glycoprotein. The SARS-CoV-2-S spike glycoprotein is a spike protein of the coronavirus SARS-CoV-2 that recently caused the worldwide spread of COVID-19 disease. This study is dedicated to the development of an electrochemical determination method based on the application of molecularly imprinted polymer technology. The electrochemical sensor was designed by molecular imprinting of polypyrrole (Ppy) with SARS-CoV-2-S spike glycoprotein (MIP-Ppy). The electrochemical sensors with MIP-Ppy and with polypyrrole without imprints (NIP-Ppy) layers were electrochemically deposited on a platinum electrode surface by a sequence of potential pulses. The performance of polymer layers was evaluated by pulsed amperometric detection. According to the obtained results, a sensor based on MIP-Ppy is more sensitive to the SARS-CoV-2-S spike glycoprotein than a sensor based on NIP-Ppy. Also, the results demonstrate that the MIP-Ppy layer is more selectively interacting with SARS-CoV-2-S glycoprotein than with bovine serum albumin. This proves that molecularly imprinted MIP-Ppy-based sensors can be applied for the detection of SARS-CoV-2 virus proteins.

12.
Polymers (Basel) ; 13(13)2021 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-34209068

RESUMO

Biosensors for the determination of glucose concentration have a great significance in clinical diagnosis, and in the food and pharmaceutics industries. In this research, short-chain polyaniline (PANI) and polypyrrole (Ppy)-based nanocomposites with glucose oxidase (GOx) and 6 nm diameter AuNPs (AuNPs(6 nm)) were deposited on the graphite rod (GR) electrode followed by the immobilization of GOx. Optimal conditions for the modification of GR electrodes by conducting polymer-based nanocomposites and GOx were elaborated. The electrodes were investigated by cyclic voltammetry and constant potential amperometry in the presence of the redox mediator phenazine methosulfate (PMS). The improved enzymatic biosensors based on GR/PANI-AuNPs(6 nm)-GOx/GOx and GR/Ppy-AuNPs(6 nm)-GOx/GOx electrodes were characterized by high sensitivity (65.4 and 55.4 µA mM-1 cm-2), low limit of detection (0.070 and 0.071 mmol L-1), wide linear range (up to 16.5 mmol L-1), good repeatability (RSD 4.67 and 5.89%), and appropriate stability (half-life period (τ1/2) was 22 and 17 days, respectively). The excellent anti-interference ability to ascorbic and uric acids and successful practical application for glucose determination in serum samples was presented for GR/PANI-AuNPs(6 nm)-GOx/GOx electrode.

13.
Int J Mol Sci ; 22(9)2021 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-34068596

RESUMO

The review focuses on the overview of electrochemical sensors based on molecularly imprinted polymers (MIPs) for the determination of uric acid. The importance of robust and precise determination of uric acid is highlighted, a short description of the principles of molecular imprinting technology is presented, and advantages over the others affinity-based analytical methods are discussed. The review is mainly concerned with the electro-analytical methods like cyclic voltammetry, electrochemical impedance spectroscopy, amperometry, etc. Moreover, there are some scattered notes to the other electrochemistry-related analytical methods, which are capable of providing additional information and to solve some challenges that are not achievable using standard electrochemical methods. The significance of these overviewed methods is highlighted. The overview of the research that is employing MIPs imprinted with uric acid is mainly targeted to address these topics: (i) type of polymers, which are used to design uric acid imprint structures; (ii) types of working electrodes and/or other parts of signal transducing systems applied for the registration of analytical signal; (iii) the description of the uric acid extraction procedures applied for the design of final MIP-structure; (iv) advantages and disadvantages of electrochemical methods and other signal transducing methods used for the registration of the analytical signal; (vi) overview of types of interfering molecules, which were analyzed to evaluate the selectivity; (vi) comparison of analytical characteristics such as linear range, limits of detection and quantification, reusability, reproducibility, repeatability, and stability. Some insights in future development of uric acid sensors are discussed in this review.


Assuntos
Técnicas Biossensoriais , Eletroquímica , Polímeros Molecularmente Impressos/química , Ácido Úrico/isolamento & purificação , Humanos , Ácido Úrico/química
14.
Sensors (Basel) ; 22(1)2021 Dec 29.
Artigo em Inglês | MEDLINE | ID: mdl-35009774

RESUMO

Polypyrrole (Ppy) and poly(methylene blue) (PMB) heterostructure (Ppy-PMB) was electrochemically formed on the indium tin oxide (ITO) coated glass slides, which served as working electrodes. For electropolymerization, a solution containing pyrrole, methylene blue, and a saccharide (lactose, sucrose, or heparin) that served as dopant was used. The aim of this study was to compare the effect of the saccharides (lactose, sucrose, and heparin) on the electrochromic properties of the Ppy-PMB layer. AFM and SEM have been used for the analysis of the surface dominant features of the Ppy-PMB layers. From these images, it was concluded that the saccharides used in this study have a moderate effect on the surface morphology. Electrochromic properties were analyzed with respect to the changes of absorbance of the layer at two wavelengths (668 nm and 750 nm) by changing the pH of the surrounding solution and the potential between +0.8 V and -0.8 V. It was demonstrated that the highest absorbance changes are characteristic for all layers in the acidic media. Meanwhile, the absorbance changes of the layers were decreased in the more alkaline media. It was determined that the Ppy-PMB layers with heparin as a dopant were more mechanically stable in comparison to the layers doped with lactose and sucrose. Therefore, the Ppy-PMB layer doped with heparin was selected for the further experiment and it was applied in the design of electrochromic sensors for the determination of three xanthine derivatives: caffeine, theobromine, and theophylline. A linear relationship of ΔA (∆A = A+0.8V - A-0.8V) vs. concentration was determined for all three xanthine derivatives studied. The largest change in optical absorption was observed in the case of theophylline determination.


Assuntos
Polímeros , Pirróis , Azul de Metileno , Polissacarídeos
15.
Polymers (Basel) ; 12(12)2020 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-33348805

RESUMO

Nanocomposites based on two conducting polymers, polyaniline (PANI) and polypyrrole (Ppy), with embedded glucose oxidase (GOx) and 6 nm size gold nanoparticles (AuNPs(6nm)) or gold-nanoclusters formed from chloroaurate ions (AuCl4-), were synthesized by enzyme-assisted polymerization. Charge (electron) transfer in systems based on PANI/AuNPs(6nm)-GOx, PANI/AuNPs(AuCl4-)-GOx, Ppy/AuNPs(6nm)-GOx and Ppy/AuNPs(AuCl4-)-GOx nanocomposites was investigated. Cyclic voltammetry (CV)-based investigations showed that the reported polymer nanocomposites are able to facilitate electron transfer from enzyme to the graphite rod (GR) electrode. Significantly higher anodic current and well-defined red-ox peaks were observed at a scan rate of 0.10 V s-1. Logarithmic function of anodic current (log Ipa), which was determined by CV-based experiments performed with glucose, was proportional to the logarithmic function of a scan rate (log v) in the range of 0.699-2.48 mV s-1, and it indicates that diffusion-controlled electrochemical processes were limiting the kinetics of the analytical signal. The most efficient nanocomposite structure for the design of the reported glucose biosensor was based on two-day formed Ppy/AuNPs(AuCl4-)-GOx nanocomposites. GR/Ppy/AuNPs(AuCl4-)-GOx was characterized by the linear dependence of the analytical signal on glucose concentration in the range from 0.1 to 0.70 mmol L-1, the sensitivity of 4.31 mA mM cm-2, the limit of detection of 0.10 mmol L-1 and the half-life period of 19 days.

16.
Talanta ; 200: 378-386, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31036199

RESUMO

Zinc oxide (ZnO) based nanostructures owing unique physical properties - high photoluminescence, biocompatibility and other characteristics, therefore, they attract attention as building blocks suitable for biosensor development. In this research as a target we have used human leukemic cell line IM9 (IM9). IM9 was derived from the patient with a multiple myeloma and expressed cluster of differentiation proteins СD19 on the surface of 85-95% here investigated cancer cells. As a control sample healthy human's peripheral blood mononuclear cells (PBMC) were used and the expression of CD19 protein was found only in 5-9% of these cells. Two types of antibodies labeled by fluorescein isothiocyanate (FITC) were used for the labeling of human leukemic cells: FITC-conjugated mouse antibodies against Human CD19 protein (anti-CD19-FITC*) and FITC-conjugated mouse antibodies against Human IgG1 protein (anti-IgG1-FITC*). In order to demonstrate the applicability of zinc oxide nanorods (ZnO-NRs) based platforms three types of ZnO-NRs-based structures were investigated: (i) ZnO-NRs modified by anti-CD19-FITC*; (ii) ZnO-NRs modified by IM9 cells, which were pre-incubated with anti-CD19-FITC*; (iii) ZnO-NRs modified by PBMC cells, which were pre-incubated with anti-CD19-FITC*. It was demonstrated that IM9 cells after specific interaction with anti-CD19-FITC* bind to ZnO-NRs (ZnO-NRs/IM9 +anti-CD19-FITC*) and photoluminescence based signal significantly increase in comparison with that observed in control samples, which contained PBMC cells incubated with anti-CD19-FITC* (ZnO-NRs/PBMC+anti-CD19-FITC*). The photoluminescence results are in good correlation with the data obtained by flow cytometry. This study illustrate that ZnO-NRs exhibit a photoluminescence signal suitable for the determination of anti-CD19-FITC* labeled IM9 cell line at concentrations - from 10 till 500 cells adsorbed per 1 mm2 of ZnO-NRs platform.


Assuntos
Linfócitos B/patologia , Separação Celular , Imunoensaio/métodos , Leucemia/patologia , Nanotubos/química , Óxido de Zinco/química , Biomarcadores/sangue , Células Cultivadas , Citometria de Fluxo , Humanos , Óxido de Zinco/síntese química
17.
Nanomaterials (Basel) ; 9(5)2019 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-31137827

RESUMO

Polyaniline (PANI), polypyrrole (Ppy), and polythiophene (PTh) composite nanoparticles with embedded glucose oxidase (GOx) were formed by enzymatic polymerization of corresponding monomers (aniline, pyrrole, and thiophene). The influence of monomers concentration, the pH of solution, and the ratio of enzyme/substrate on the formation of PANI/GOx, Ppy/GOx, and PTh/GOx composite nanoparticles were spectrophotometrically investigated. The highest formation rate of PANI-, Ppy-, and PTh-based nanoparticles with embedded GOx was observed in the sodium acetate buffer solution, pH 6.0. The increase of optical absorbance at λmax = 440 nm, λmax = 460 nm, and λmax = 450 nm was exploited for the monitoring of PANI/GOx, Ppy/GOx and PTh/GOx formation, respectively. It was determined that the highest polymerization rate of PANI/GOx, Ppy/GOx, and PTh/GOx composite nanoparticles was achieved in solution containing 0.75 mg mL-1 of GOx and 0.05 mol L-1 of glucose. The influence of the enzymatic polymerization duration on the formation of PANI/GOx and Ppy/GOx composite nanoparticles was spectrophotometrically investigated. The most optimal duration for the enzymatic synthesis of PANI/GOx and Ppy/GOx composite nanoparticles was in the range of 48-96 h. It was determined that the diameter of formed PANI/GOx and Ppy/GOx composite nanoparticles depends on the duration of polymerization using dynamic light scattering technique (DLS), and it was in the range of 41-167 nm and 65-122 nm, when polymerization lasted from 16 to 120 h.

18.
Polymers (Basel) ; 11(2)2019 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-30960361

RESUMO

Several types of polyaniline (PANI) and polypyrrole (Ppy) nanocomposites with embedded glucose oxidase (GOx) and gold nanoparticles (AuNPs) were formed by enzymatic polymerization of corresponding monomers (aniline and pyrrole) in the presence of 6 and 13 nm diameter colloidal gold nanoparticles (AuNPs(6nm) or AuNPs(13nm), respectively) or chloroaurate ions (AuCl4-). Glucose oxidase in the presence of glucose generated H2O2, which acted as initiator of polymerization reaction. The influence of polymerization bulk composition and pH on the formation of PANI- and Ppy-based nanocomposites was investigated spectrophotometrically. The highest formation rate of PANI- and Ppy-based nanocomposites with embedded glucose oxidase and gold nanoparticles (PANI/AuNPs-GOx and Ppy/AuNPs-GOx, respectively) was observed in the solution of sodium acetate buffer, pH 6.0. It was determined that the presence of AuNPs or AuCl4- ions facilitate enzymatic polymerization of aniline and pyrrole.

19.
Crit Rev Anal Chem ; 49(3): 195-208, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30285474

RESUMO

Electrochromic sensors offer multi-mode registration of analytical signal based on combination of electrochemical and optical techniques. This emerging direction of analytical chemistry is relatively new; therefore, it has very high potential for various applications in chemical and biochemical analysis. Properties of sensors based on various electrochromic materials such as polymers, polymer derivatives, polymer composites, metal oxides, metal oxide complexes, phthalocyanines, porphyrins, and dyes are critically overviewed, evaluated, and compared. The most promising directions in analytical application of electrochromic polymers are highlighted.


Assuntos
Complexos de Coordenação/análise , Técnicas Eletroquímicas , Metais/análise , Óxidos/análise , Polímeros/análise , Corantes/análise , Indóis/análise , Isoindóis , Porfirinas/análise
20.
Colloids Surf B Biointerfaces ; 175: 671-679, 2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30590328

RESUMO

In this research we report the biological synthesis of electrically conducting polymer - Polypyrrole (Ppy). Cell-assisted enzymatic polymerization/oligomerization of Ppy was achieved using whole cell culture and cell-free crude enzyme extract from two white-rot fungal cultures. The selected fungal strains belong to Trametes spp., known laccase producers, commonly applied in bioremediation and bioelectrochemical fields. The biocatalytic reaction was initiated in situ through the copper-containing enzymes biosynthesized within the cell cultures under submerged aerobe cultivation conditions. The procedure was inspired by successful reports of laccase-catalyzed pyrrole polymerization. The usage of whole culture and/or crude enzyme extract has the advantage of overcoming enzyme purification and minimizing the liability of enzyme inactivation through improved stability of enzymes in their natural environment. Spectral and electrochemical techniques (UV-vis spectroscopy, infrared spectroscopy; cyclic voltammetry (CV)) and pH measurements provided insight into the evolution of pyrrole polymerization/oligomerization and the electrochemical features of the final product. Microscopy techniques (optical microscopy and scanning electron microscopy (SEM)) were primary tools for visualization of the formed Ppy particles. The relevance of our research is twofold: Ppy prepared in crude enzyme extract results in enzyme encapsulated within Ppy and/or Ppy-modified fungal cells can be formed when polymerization occurs in whole cell culture. The route of biocatalysis can be chosen according to the desired bioelectrochemical application. The reported study focuses on the improvement of charge transfer through the fungal cell membrane and/or cell wall by modification of the fungal cells with conducting polymer - polypyrrole.


Assuntos
Proteínas Fúngicas/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Polímeros/metabolismo , Pirróis/metabolismo , Trametes/metabolismo , Benzotiazóis/metabolismo , Benzotiazóis/farmacologia , Biocatálise , Parede Celular/efeitos dos fármacos , Parede Celular/metabolismo , Meios de Cultura/química , Meios de Cultura/farmacologia , Condutividade Elétrica , Técnicas Eletroquímicas , Fermentação/efeitos dos fármacos , Guaiacol/metabolismo , Guaiacol/farmacologia , Hidrazonas/metabolismo , Hidrazonas/farmacologia , Concentração de Íons de Hidrogênio , Polímeros/química , Pirróis/química , Ácidos Sulfônicos/metabolismo , Ácidos Sulfônicos/farmacologia , Trametes/efeitos dos fármacos
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