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1.
Biosensors (Basel) ; 13(6)2023 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-37367006

RESUMO

Bacterial infections resulting from foodborne pathogenic bacteria cause millions of infections that greatly threaten human health and are one of the leading causes of mortality around the world. To counter this, the early, rapid, and accurate detection of bacterial infections is very important to address serious health issue concerns. We, therefore, present an electrochemical biosensor based on aptamers that selectively bind with the DNA of specific bacteria for the accurate and rapid detection of various foodborne bacteria for the selective determination of bacterial infection types. Different aptamers were synthesized and immobilized on Au electrodes for selective bindings of different types of bacterial DNA (Escherichia coli, Salmonella enterica, and Staphylococcus aureus) for the accurate detection and quantification of bacterial concentrations from 101 to 107 CFU/mL without using any labeling methods. Under optimized conditions, the sensor showed a good response to the various concentrations of bacteria, and a robust calibration curve was obtained. The sensor could detect the bacterial concentration at meager quantities and possessed an LOD of 4.2 × 101, 6.1 × 101, and 4.4 × 101 CFU/mL for S. Typhimurium, E. Coli, and S. aureus, respectively, with a linear range from 100 to 104 CFU/mL for the total bacteria probe and 100 to 103 CFU/mL for individual probes, respectively. The proposed biosensor is simple and rapid and has shown a good response to bacterial DNA detections and thus can be applied in clinical applications and food safety monitoring.


Assuntos
Aptâmeros de Nucleotídeos , Infecções Bacterianas , Técnicas Biossensoriais , Humanos , Escherichia coli/genética , DNA Bacteriano , Staphylococcus aureus , Sistemas Automatizados de Assistência Junto ao Leito , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Limite de Detecção , Técnicas Eletroquímicas/métodos
2.
Biosensors (Basel) ; 13(1)2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36671926

RESUMO

Technological advancements in the healthcare sector have pushed for improved sensors and devices for disease diagnosis and treatment. Recently, with the discovery of numerous biomarkers for various specific physiological conditions, early disease screening has become a possibility. Biomarkers are the body's early warning systems, which are indicators of a biological state that provides a standardized and precise way of evaluating the progression of disease or infection. Owing to the extremely low concentrations of various biomarkers in bodily fluids, signal amplification strategies have become crucial for the detection of biomarkers. Metal nanoparticles are commonly applied on 2D platforms to anchor antibodies and enhance the signals for electrochemical biomarker detection. In this context, this review will discuss the recent trends and advances in metal nanoparticle decorated 2D materials for electrochemical biomarker detection. The prospects, advantages, and limitations of this strategy also will be discussed in the concluding section of this review.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Técnicas Eletroquímicas , Biomarcadores , Anticorpos , Ouro , Imunoensaio
3.
Bioelectrochemistry ; 150: 108347, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36549174

RESUMO

Microelectrode-based cell chip studies for cellular responses often require improved adhesion and growth conditions for efficient cellular diagnosis and high throughput screening in drug discovery. Cell-chip studies are often performed on gold electrodes due to their biocompatibility, and stability, but the electrode-electrolyte interfacial capacitance is the main drawback to the overall sensitivity of the detection system. Thus, here, we developed reduced graphene oxide-polyaniline-modified gold microelectrodes for real-time impedance-based monitoring of human gastric adenocarcinoma cancer (MKN-1) cells. The impedance characterization on modified electrodes showed 28-fold enhanced conductivity than the bare electrodes, and the spectra were modeled with proper equivalent circuits to extrapolate the values of circuit elements. The impedance of both time-and frequency-dependent measurements of cell-covered modified electrodes with equivalent model circuits was analyzed to achieve cellular behavior, such as adhesion, spreading, proliferation, and influence of anti-cancer agents. The normalized impedance at 41.5 kHz (|Z|norm 41 kHz) was selected to monitor the cell growth analysis, which was found linear with the proliferation of adherent cells along with the influence of the anticancer drug agent on the MKN-1 cells. The synergistic effects and biocompatible nature of PANI-RGO modifications improved the overall sensitivity for the cell-growth studies of MKN-1 cells.


Assuntos
Grafite , Neoplasias , Humanos , Microeletrodos , Impedância Elétrica , Ouro
4.
Anal Chem ; 94(49): 17020-17030, 2022 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-36414244

RESUMO

A novel electrochemical sensor was constructed based on an enzyme-mediated physiological reaction between neurotransmitter serotonin per-oxidation to reconstruct dual-molecule 4,4'-dimeric-serotonin self-assembled derivative, and the potential biomedical application of the multi-functional nano-platform was explored. Serotonin accelerated the catalytic activity to form a dual molecule at the C4 position and created phenolic radical-radical coupling intermediates in a peroxidase reaction system. Here, 4,4' dimeric-serotonin possessed the capability to recognize intermolecular interactions between amine groups. The excellent quenching effects on top of the gold surface electrode system archive logically inexpensive and straightforward analytical demands. In biochemical sensing analysis, the serotonin dimerization concept demonstrated a robust, low-cost, and highly sensitive immunosensor, presenting the potential of quantifying serotonin at point-of-care (POC) testing. The high-specificity serotonin electrochemical sensor had a limit of detection (LOD) of 0.9 nM in phosphate buffer and 1.4 nM in human serum samples and a linear range of 10 to 400 with a sensitivity of 2.0 × 10-2 nM. The bivalent 4,4'-dimer-serotonin interaction strategy provides a promising platform for serotonin biosensing with high specificity, sensitivity, selectivity, stability, and reproducibility. The self-assembling gold surface electrochemical system presents a new analytical method for explicitly detecting tiny neurotransmitter-responsive serotonin neuromolecules.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Humanos , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , Serotonina/análise , Reprodutibilidade dos Testes , Imunoensaio/métodos , Ouro/química , Eletrodos , Limite de Detecção , Polímeros , Neurotransmissores/análise , Nanopartículas Metálicas/química
5.
Anal Bioanal Chem ; 414(10): 3205-3217, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-34617153

RESUMO

Sensitive and selective determination of protein biomarkers with high accuracy often remains a great challenge due to their existence in the human body at an exceptionally low concentration level. Therefore, sensing mechanisms that are easy to use, simple, and capable of accurate quantification of analyte are still in development to detect biomarkers at a low concentration level. To meet this end, we demonstrated a methodology to detect thrombin in serum at low concentration levels using polypyrrole (PPy)-palladium (Pd)nanoparticle-based hybrid transducers using liposomes encapsulated redox marker as a label. The morphology of Ppy-Pd composites was characterized by scanning electron microscopy, and the hybrid structure provided excellent binding and detection platform for thrombin detection in both buffer and serum solutions. For quantitative measurement of thrombin in PBS and serum, the change in current was monitored using differential pulse voltammetry, and the calculated limit of quantification (LOQ) and limit of detection (LOD) for the linear segment (0.1-1000 nM of thrombin) were 1.1 pM and 0.3 pM, in serum, respectively. The sensors also exhibited good stability and excellent selectivity towards the detection of thrombin, and thus make it a strong candidate for adopting its sensing applications in biomarker detection technologies.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanocompostos , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Humanos , Limite de Detecção , Lipossomos , Paládio/química , Polímeros/química , Pirróis/química , Trombina/química
6.
Biosens Bioelectron ; 191: 113447, 2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34175648

RESUMO

Chemically modulating monoamine neurotransmitter serotonin undergoes a physiological reaction of enzyme intermediated peroxidation to reconstruct dimeric self-assembled complex. A standard bivalent ligand approach dimeric serotonin increases structural and functional scaffolding with recognition-binding sites that are fundamentally more friendly than monovalent binding sites. Dimerization reaction accelerates the catalytic activity of one-electron oxidation at the C(4) position of serotonin to generate dual phenolic radicals in the presence of horseradish (HRP) and hydrogen peroxide (H2O2). Herein, we suggest the dimeric serotonin-based colorimetric assay, which presents a new rapid, sensitive, selective, and quantitative visualization. The dimeric serotonin possesses the capability to recognize intermolecular interaction units that cause aggregation scaffold of gold nanoparticles (GNPs), providing inexpensive and straightforward analytical needs. As a proof of visual and spectral analysis, peroxidative dimeric serotonin demonstrated sensitive and robust results. The calorimetric method enables highly sensitive detection of serotonin in phosphate buffer, and in human serum samples at nanomolar levels with a LOD of 2.6 nM and 2.81 nM, respectively, and the sensor possesses a dynamic range of 100-300 nM in buffer condition. Also, as proof of concept, visible color imaging of immunosensors which is appropriate for fast visible testing at detection limits as low as 2.90 nM concentration.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Colorimetria , Ouro , Humanos , Peróxido de Hidrogênio , Imunoensaio , Ligantes , Limite de Detecção , Serotonina
7.
Nanomaterials (Basel) ; 10(9)2020 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-32825146

RESUMO

Hydrogen peroxide (H2O2) is a partially reduced metabolite of oxygen that exerts a diverse array of physiological and pathological activities in living organisms. Therefore, the accurate quantitative determination of H2O2 is crucial in clinical diagnostics, the food industry, and environmental monitoring. Herein we report the electrosynthesis of silver nanoflowers (AgNFs) on indium tin oxide (ITO) electrodes for direct electron transfer of hemoglobin (Hb) toward the selective quantification of H2O2. After well-ordered and fully-grown AgNFs were created on an ITO substrate by electrodeposition, their morphological and optical properties were analyzed with scanning electron microscopy and UV-Vis spectroscopy. Hb was immobilized on 3-mercaptopropionic acid-coated AgNFs through carbodiimide cross-linking to form an Hb/AgNF/ITO biosensor. Electrochemical measurement and analysis demonstrated that Hb retained its direct electron transfer and electrocatalytic properties and acted as a H2O2 sensor with a detection limit of 0.12 µM and a linear detection range of 0.2 to 3.4 mM in phosphate-buffered saline (PBS). The sensitivity, detection limit, and detection range of the Hb/AgNF/ITO biosensor toward detection H2O2 in human serum was also found to be 0.730 mA mM-1 cm-2, 90 µM, and 0.2 to 2.6 mM, indicating the clinical application for the H2O2 detection of the Hb/AgNF/ITO biosensor. Moreover, interference experiments revealed that the Hb/AgNF/ITO sensor displayed excellent selectivity for H2O2.

8.
Biosens Bioelectron ; 164: 112272, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32553348

RESUMO

Highly porous laser-induced graphene (LIG) is easily generated in complex electrode configurations such as interdigitated electrodes (IDEs). Here, we demonstrate that their superior capacitive response at low frequencies can be exploited in affinity biosensors using thrombin aptamers as model biorecognition elements. Of specific interest was the effect of electrode surface area on capacitance detection, and the comparison between a label-free format and enhancement strategies afforded by carboxy group bearing polymeric nanoparticles or liposomes. Electrochemical impedance spectroscopy (EIS) was used to investigate the LIG performance and optimize the biosensor design. Interestingly, the label-free strategy performed extremely well and additional labels decreased the limit of detection or increased the sensitivity only minimally. It is assumed that the highly porous nature of the LIG structures dominates the capacitive response so that labels removed from the surface have only limited influence Also, while slight performance changes can be observed for smaller vs. larger electrode structures, the performance of a LIG IDE is reasonably independent of its size. In the end, a dynamic range of 5 orders of magnitude was obtained (0.01 nM-1000 nM) with a limit of detection as low as 0.12 pM. When measured in serum, this increased to 1.3 pM. The good reproducibility (relative standard deviation (RSD), 4.90%) and repeatability (RSD, 2.59%) and good long-term stability (>7 weeks at 4 °C) prove that a LIG-based capacitance sensor is an excellent choice for affinity-based biosensor. The ease-of-production, the simplicity of modification and the superior performance even in a label-free format indicate that LIG-based biosensors should be considered in point-of-care diagnostics in the future.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Grafite , Eletrodos , Lasers , Reprodutibilidade dos Testes
9.
Anal Chem ; 91(9): 5841-5849, 2019 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-30938982

RESUMO

Infectious pancreatic necrosis virus (IPNV) has been identified as a viral pathogen for many fish diseases that have become a huge hurdle for the growing fishing industry. Thus, in this work, we report a label-free impedance biosensor to quantify IPNV in real fish samples at point-of-care (POC) level. High specificity IPNV sensor with a detection limit of 2.69 TCID50/mL was achieved by conjugating IPNV antibodies to portable Au disk electrode chips using human heavy chain apoferritin (H-AFN) nanoprobes as a binding agent. H-AFN probes were bioengineered through PCR by incorporating pET-28b(+) resulting in 24 subunits of 6 × his-tag and protein-G units on its outer surface to increase the sensitivity of the IPNV detection. The biosensor surface modifications were characterized by differential pulse voltammetry (DPV) and EIS methods for each modification step. The proposed nanoprobe based sensor showed three-fold enhancement in charge transfer resistance toward IPNV detection in comparison with the traditional linker approach when measured in a group of similar virus molecules. The portable sensor exhibited a linear range of 100-10000 TCID50/mL and sensitivity of 5.40 × 10-4 TCID50/mL in real-fish samples. The performance of the proposed IPNV sensor was fully validated using an enzyme-linked immunosorbent assay (ELISA) technique with a sensitivity of 3.02 × 10-4 TCID50/mL. Results from H-AFN nanoprobe based IPNV sensor indicated high selectivity, sensitivity, and stability could be a promising platform for the detection of similar fish viruses and other biological molecules of interest.


Assuntos
Apoferritinas/química , Técnicas Biossensoriais/métodos , Infecções por Birnaviridae/veterinária , Doenças dos Peixes/diagnóstico , Peixes/virologia , Vírus da Necrose Pancreática Infecciosa/isolamento & purificação , Nanopartículas/química , Animais , Bioengenharia , Infecções por Birnaviridae/diagnóstico , Infecções por Birnaviridae/virologia , Doenças dos Peixes/virologia
10.
Bioelectrochemistry ; 128: 165-174, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31004910

RESUMO

Most clinical tests for biomarker detection require the support of a laboratory, and the results are usually slow, less sensitive, and lack the possibility for Point-of-Care (PoC) testing. Further, with the increasing demand for sensitive, portable, rapid, and low-cost devices for clinical PoC applications, innovative methods are crucial. Thus, we report on utilizing nanostructured gold-platinum (Au-Pt) hybrid electrodes as a PoC device for highly sensitive and selective PTH detection in human serum samples. The method employs the immobilization of 3-mercaptopropionic acid to Au and subsequent activation of the carboxyl groups to enable anti-PTH antibody immobilization. Serum PTH was detected by monitoring the changes in electrochemical properties (ΔRct and Δi) of the sensor using electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV) against a standard hexacyanoferrate (II/III) probe. Changes in relative response percentage (RR%) in electrochemical properties due to increased PTH concentrations in serum were observed with EIS and DPV. The biosensor exhibited a low detection limit of 0.36 pg.mL-1 (EIS) and 0.59 pg.mL-1 (DPV) in serum with a linear range of 1 to 100,000 pg.mL-1. Further, to validate the accuracy of the proposed method, clinical samples (n = 20) were examined using the EIS method and compared to an established commercial test.


Assuntos
Eletrodos , Ouro/química , Nanoestruturas , Hormônio Paratireóideo/sangue , Platina/química , Estudos de Casos e Controles , Espectroscopia Dielétrica , Ensaio de Imunoadsorção Enzimática , Humanos , Hiperparatireoidismo/sangue , Limite de Detecção , Sistemas Automatizados de Assistência Junto ao Leito , Reprodutibilidade dos Testes
11.
Sensors (Basel) ; 18(5)2018 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-29695134

RESUMO

Aflatoxin B1 (AFB1) is produced by the Aspergillus flavus and Aspergillus parasiticus group of fungi which is most hepatotoxic and hepatocarcinogenic and occurs as a contaminant in a variety of foods. AFB1 is mutagenic, teratogenic, and causes immunosuppression in animals and is mostly found in peanuts, corn, and food grains. Therefore, novel methodologies of sensitive and expedient strategy are often required to detect mycotoxins at the lowest level. Herein, we report an electrochemical impedance sensor that selectively detects AFB1 at the lowest level by utilizing polyaniline nanofibers (PANI) coated with gold (Au) nanoparticles composite based indium tin oxide (ITO) disk electrodes. The Au-PANI nanocomposites were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) spectroscopy, and electrochemical impedance spectroscopy (EIS). The composite electrode exhibited a 14-fold decrement in |Z|1Hz in comparison with the bare electrode. The Au-PANI acted as an effective sensing platform having high surface area, electrochemical conductivity, and biocompatibility which enabled greater loading deposits of capture antibodies. As a result, the presence of AFB1 was screened with high sensitivity and stability by monitoring the changes in impedance magnitude (|Z|) in the presence of a standard iron probe which was target specific and proportional to logarithmic AFB1 concentrations (CAFB1). The sensor exhibits a linear range 0.1 to 100 ng/mL with a detection limit (3) of 0.05 ng/mL and possesses good reproducibility and high selectivity against another fungal mycotoxin, Ochratoxin A (OTA). With regard to the practicability, the proposed sensor was successfully applied to spiked corn samples and proved excellent potential for AFB1 detection and development of point-of-care (POC) disease sensing applications.


Assuntos
Eletrodos , Aflatoxina B1 , Compostos de Anilina , Técnicas Biossensoriais , Impedância Elétrica , Técnicas Eletroquímicas , Ouro , Nanopartículas Metálicas , Nanofibras , Reprodutibilidade dos Testes
12.
Bioelectrochemistry ; 122: 93-102, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29602041

RESUMO

Highly sensitive and selective biosensors for accurate determination of specific protein biomarkers at low levels in serum are a prerequisite for the present healthcare systems. Therefore, here we developed a label-free impedimetric method for tumor necrosis factor-alpha (TNF-α) detection using reduced graphene oxide (RGO) with gold nanoparticles (AuNP) on an indium tin oxide (ITO) microdisk electrodes. The detection mechanism relies on resistance change occurs due to [Fe(CN)6]3-/4- redox probe movement towards the conductive channels of the AuNP-RGO films gated by the recognition of the target biomarker by its anti-TNF-α antibody. The conductivity of the AuNP-RGO structures enhanced by 10-fold in comparison with bare electrode chips. The observed resistance changes at 2Hz (ΔR at 2Hz) enabled the quantification of various concentrations of TNF-α in human serum (CHS-TNF-α). The antibody-adsorbed electrode showed a good increment of resistance change (ΔR) with additions of antigen concentration. The sensor possesses a linear range of 1-1000pg/ml had a detection limit of 0.67pg/ml (38.51fM) and 0.78pg/ml (44.83fM) in PBS and human serum, respectively.


Assuntos
Anticorpos Imobilizados/química , Técnicas Biossensoriais/métodos , Fator de Necrose Tumoral alfa/sangue , Técnicas Eletroquímicas/métodos , Ouro/química , Grafite/química , Humanos , Imunoensaio/métodos , Limite de Detecção , Nanopartículas Metálicas/química , Nanopartículas Metálicas/ultraestrutura , Oxirredução
13.
Materials (Basel) ; 10(7)2017 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-28773163

RESUMO

In the present study, we propose a method for bio-nano hybrid formation by coupling a redox metalloprotein, Azurin, with CdSe-ZnS quantum dot for the development of a nanoscale resistive memory device. The covalent interaction between the two nanomaterials enables a strong and effective binding to form an azurin/CdSe-ZnS hybrid, and also enabled better controllability to couple with electrodes to examine the memory function properties. Morphological and optical properties were performed to confirm both hybrid formations and also their individual components. Current-Voltage (I-V) measurements on the hybrid nanostructures exhibited bistable current levels towards the memory function device, that and those characteristics were unnoticeable on individual nanomaterials. The hybrids showed good retention characteristics with high stability and durability, which is a promising feature for future nanoscale memory devices.

14.
Biosens Bioelectron ; 90: 23-30, 2017 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-27871046

RESUMO

In the present study, we developed a biohybrid material composed of recombinant azurin and CdSe-ZnS quantum dot to perform as a resistive random access memory (ReRAM) device. Site specific amino acid sequences were introduced in azurin to bind with the surface of CdSe-ZnS nanoparticle allowing the formation of a hybrid and voltage-driven switching enabled to develop a resistive random access memory (ReRAM) device. The analytical measurements confirmed that the azurin and CdSe-ZnS nanoparticles were well conjugated and formed into a single hybrid. Further, reversible, bistable switching along with repeatable writing-reading-erasing processes on individual azurin/CdSe-ZnS hybrid at nanoscale was achieved on the hybrid device. The device was programmed tested for 50 cycles with an ON/OFF ratio and measured to be of three orders of magnitude. The developed device shown good stability and repeatability and operates at low voltages thus makes it promising candidate for future memory device applications.


Assuntos
Azurina/química , Técnicas Biossensoriais/instrumentação , Nanopartículas/química , Nanotecnologia/instrumentação , Sequência de Aminoácidos/genética , Azurina/genética , Técnicas Biossensoriais/métodos , Compostos de Cádmio/química , Nanotecnologia/métodos , Pontos Quânticos/química , Compostos de Selênio/química , Processamento de Sinais Assistido por Computador , Zinco/química
15.
Biosens Bioelectron ; 80: 307-314, 2016 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-26852199

RESUMO

Sensitive and selective determination of protein biomarkers remains a significant challenge due to the existence of various biomarkers in human body at a low concentration level. Therefore, new technologies were incessantly steered to detect tiny biomarkers at a low concentration level, yet, it is difficult to develop reliable, stable and sensitive detection methods for disease diagnostics. Therefore, the present study demonstrates a methodology to detect insulin in serum at low levels based on Ag nanoflower (AgNF) decorated reduced graphene oxide (rGO) modified micro-disk electrode arrays (MDEAs). The morphology of AgNF-rGO composite was characterized by scanning electron microscopy, the structure was analyzed using X-ray diffraction patterns and Raman spectra. The hybrid interface exhibited enhanced electrical conductivity when compared with its individual elements and had improved capturing ability for antibody-antigen binding towards insulin detection. In order to measure quantitatively the insulin concentration in PBS and human serum, the change in impedance (ΔZ) from electrochemical impedance spectroscopy was analyzed for various concentrations of insulin in [Fe(CN)6](3-/4-) redox couple. The electrode with adsorbed antibodies showed an increase in ΔZ for the addition of antigen concentrations over a working range of 1-1000 ng mL(-1). The detection limits were 50 and 70 pg mL(-1) in PBS and human serum, respectively.


Assuntos
Técnicas Biossensoriais/instrumentação , Espectroscopia Dielétrica/instrumentação , Grafite/química , Insulina/sangue , Nanoestruturas/química , Prata/química , Anticorpos Imobilizados/química , Eletrodos , Humanos , Limite de Detecção , Modelos Moleculares , Oxirredução , Óxidos/química
16.
Sensors (Basel) ; 16(1)2016 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-26784202

RESUMO

Insulin is a key regulator in glucose homeostasis and its deficiency or alternations in the human body causes various types of diabetic disorders. In this paper, we present the development of a reduced graphene oxide (rGO) modified interdigitated chain electrode (ICE) for direct capacitive detection of insulin. The impedance properties of rGO-ICE were characterized by equivalent circuit modeling. After an electrochemical deposition of rGO on ICE, the electrode was modified with self-assembled monolayers and insulin antibodies in order to achieve insulin binding reactions. The impedance spectra and capacitances were measured with respect to the concentrations of insulin and the capacitance change (ΔC) was analyzed to quantify insulin concentration. The antibody immobilized electrode showed an increment of ΔC according to the insulin concentration in human serum ranging from 1 ng/mL to 10 µg/mL. The proposed sensor is feasible for label-free and real-time measuring of the biomarker and for point-of-care diagnosis.


Assuntos
Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Grafite/química , Insulina/sangue , Óxidos/química , Espectroscopia Dielétrica , Capacitância Elétrica , Eletrodos , Desenho de Equipamento , Humanos , Insulina/química
17.
Bioelectrochemistry ; 107: 37-44, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26523504

RESUMO

For label-free and direct detection of C-reactive protein (CRP), an impedimetric sensor based on an indium tin oxide (ITO) electrode array functionalized with reduced graphene oxide-nanoparticle (rGO-NP) hybrid was fabricated and evaluated. Analytical measurements were performed to examine the properties of rGO-NP-modified ITO microelectrodes and to determine the influence upon sensory performance of using nanostructures modified for antibody immobilization and for recognition of CRP binding events. Impedimetric measurements in the presence of the redox couple [Fe(CN)6](3-/4-) showed significant changes in charge transfer resistance upon binding of CRP. The impedance measurements were highly target specific, linear with logarithmic CRP concentrations in PBS and human serum across a 1 ng mL(-1) and 1000 ng mL(-1) range and associated with a detection limits of 0.06 and 0.08 ng mL(-1) respectively.


Assuntos
Técnicas Biossensoriais/métodos , Proteína C-Reativa/análise , Grafite/química , Nanopartículas/química , Compostos de Estanho/química , Anticorpos Imobilizados/química , Técnicas Biossensoriais/instrumentação , Proteína C-Reativa/imunologia , Eletroquímica , Eletrodos , Desenho de Equipamento , Propriedades de Superfície
18.
ACS Nano ; 9(7): 6675-82, 2015 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-26135474

RESUMO

RNA nanotechnology offers advantages to construct thermally and chemically stable nanoparticles with well-defined shape and structure. Here we report the development of an RNA-QD (quantum dot) chimera for resistive biomolecular memory application. Each QD holds two copies of the pRNA three-way junction (pRNA-3WJ) of the bacteriophage phi29 DNA packaging motor. The fixed quantity of two RNAs per QD was achieved by immobilizing the pRNA-3WJ with a Sephadex aptamer for resin binding. Two thiolated pRNA-3WJ serve as two feet of the chimera that stand on the gold plate. The RNA nanostructure served as both an insulator and a mediator to provide defined distance between the QD and gold. Immobilization of the chimera nanoparticle was confirmed with scanning tunneling microscopy. As revealed by scanning tunneling spectroscopy, the conjugated pRNA-3WJ-QD chimera exhibited an excellent electrical bistability signal for biomolecular memory function, demonstrating great potential for the development of resistive biomolecular memory and a nano-bio-inspired electronic device for information processing and computing.


Assuntos
Aptâmeros de Nucleotídeos/química , Nanotecnologia/métodos , Pontos Quânticos/química , RNA Viral/química
19.
J Nanosci Nanotechnol ; 15(10): 7881-5, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26726433

RESUMO

Nano or microelectrode-based cell chip for stimulating or recording neuronal signals requires better cell adhesion procedures in order to achieve efficient cell based assays for effective cellular diagnosis and for high throughput screening of drug candidates. The cells can be adhered on protein pre-coated sensing electrodes, but the electrochemical characteristics of cells are highly influenced by the electrical charge of the underlying protein interface. Thus, in this study, we report on experimental and theoretical aspects of poly-L-lysine (PLL) adsorption on transparent indium tin oxide (ITO) electrodes and the interaction between PLL and human embryonic kidney 293/GFP cells. PLL coated ITO electrodes showed a lower transfer resistance compared to bare or bovine serum albumin coated ITO electrodes. In addition, they exhibited more positive potential and higher magnitude of redox peak currents with increased immersion time of PLL solution. Finally, results of the impedance analysis showed that adhesion of cells was enhanced by PLL coating on ITO electrodes compared to bare ITO electrodes.


Assuntos
Materiais Revestidos Biocompatíveis/química , Técnicas Eletroquímicas , Polilisina/química , Soroalbumina Bovina/química , Compostos de Estanho/química , Animais , Bovinos , Adesão Celular , Eletrodos , Células HEK293 , Humanos
20.
J Nanosci Nanotechnol ; 13(9): 6424-8, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24205675

RESUMO

A protein based electrochemical sensor for the detection of hydrogen peroxide based on Myoglobin immobilized on gold nano structures patterned on Indium tin oxide electrode was developed. A uniformly distributed nanometer sized Au-array on ITO electrode surface was obtained by optimizing electro deposition conditions. The morphology of Mb molecules and Au-nanostructures on ITO was investigated by scanning electron microscopy. A Cyclic voltammetry technique was employed to study electrochemical behavior of immobilized Mb on Au/ITO electrode. From CV, a pair of quasi-reversible redox peaks of Mb obtained in 10 mM PBS buffer solution at 0.28 and 0.11 V respectively. From the electrochemical experiments, it is observed that Mb/Au/ITO electrode provides a facile electron transfer between Mb and modified ITO electrode and it also catalyzes the reduction of H2O2. A linear increase in amperometric current with increase in H2O2 concentration was also observed. The stability, reusability and selectivity of the biosensor were also evaluated. The proposed biosensor exhibits an effective and fast catalytic response to reduction of H2O2 which can be used in future biosensor applications.


Assuntos
Técnicas Eletroquímicas/métodos , Eletrodos , Ouro/química , Peróxido de Hidrogênio/análise , Mioglobina/química , Animais , Catálise , Cavalos , Microscopia Eletrônica de Varredura , Espectrofotometria Ultravioleta
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