Review Immobilization Techniques in the Fabrication of

Electroanalysis

Nitric oxide radical (NO) is a signalling molecule involved in virtually all forms of life. Its relevance has been leading to the development of different analytical methodologies to assess the temporal and spatial fluxes of NO under the complex biological milieu. Third-generation electrochemical biosensors are promising tools for in loco and in vivo NO quantification and, over the past years, heme proteins and porphyrins have been used in their design. Since there are some limitations with the biorecognition element directly adsorbed onto the electrode surface, nanomaterials (carbon nanotubes, gold nanoparticles, etc.) and polymers (cellulose, chitosan, nafion , polyacrylamide, among others) have been explored to achieve high kinetics and better biosensor performance. In this review, a broad overview of the field of electrochemical third-generation biosensors for NO electroanalysis is presented, discussing their main characteristics and aiming new outlooks and advances in this field.

Journal of Analysis and Testing

An original interfacial hybrid cross-linker molecule combined electrochemical and photochemical properties by substitution of two functional groups, pyrene, and diazirine. The first group enables anchoring by strong p-stacking interaction or by electropolymerization onto multi-walled carbon nanotubes (MWCNT). The other photoreactive group allows the covalent link with biomolecules under long-wavelength UV illumination. Diazirine was chosen due to its ability to photogenerate high reactive radicals under UV irradiation. The photoreactivity of these immobilized nanostructured conductive surfaces was tested towards the covalent attachment of tyrosinase which is well known to oxidize a large range of phenolic compounds and its yield and availability was evaluated by amperometric measurements of catechol by using molecular dioxygen. The architecture exhibiting the best analytical characteristics obtained for catechol was then chosen to detect dopamine.

Nanomaterials, 2019

Electrochemical immunosensors are antibody-based affinity biosensors with a high impact on clinical, environmental, food, and pharmaceutical analysis. In general, the analytical performance of these devices is critically determined by the materials and reagents used for their construction, signal production and amplification. Dendrimers are monodisperse and highly branched polymers with three-dimensional structures widely employed as “soft” nanomaterials in electrochemical immunosensor technology. This review provides an overview on the state-of-the-art in dendrimer-based electrochemical immunosensors, focusing on those using polyamidoamine and poly (propylene imine) dendrimers. Special emphasis is given to the most original methods recently reported for the construction of immunosensor architectures incorporating dendrimers, as well as to novel sensing approaches based on dendrimer-assisted signal enhancement strategies.

This paper reviews the engineering design of an electrochemical biosensor, particularly the main concepts of electrodes and the type of material selections, design, and fabrication method. Furthermore, the related theories and practical examples from existing literature are reviewed. Research is now directed toward the development of biosensors as important bioanalytical tools in the pharmaceutical, biotechnology, food, and other consumeroriented industries. Currently, comprehensive guidelines on the selection of electrodes for electro

Journal of Chemistry, 2015

Indinavir is a protease inhibitor antiretroviral (ARV) drug, which forms part of the highly active antiretroviral therapy during the treatment of HIV/AIDS. Indinavir undergoes first-pass metabolism through the cytochrome P450 (CYP) enzymes in the human liver, of which CYP3A4 is the most influential isoenzyme. Multidrug combination therapy and, as such, therapeutic drug monitoring (TDM) during HIV/AIDS treatment are therefore critical, to prevent adverse interactions. The conventional sensitive and specific assays available for quantifying ARV drugs, however, suffer from distinct disadvantages. In this regard, biosensors can be used to provide real time information on the metabolic profile of the drug. In this study, a biosensor with cobalt(III) sepulchrate trichloride{CoSep3+}as diffusional mediator was constructed. The biosensor platform consisted of CYP3A4 immobilized onto a gold nanoparticle (GNP) overoxidized polypyrrole (OvOxPpy) carrier matrix. The biosensor exhibited reversib...

Chemical Reviews

Bioelectrocatalysis is an interdisciplinary research field combining biocatalysis and electrocatalysis via the utilization of materials derived from biological systems as catalysts to catalyze the redox reactions occurring at an electrode. Bioelectrocatalysis synergistically couples the merits of both biocatalysis and electrocatalysis. The advantages of biocatalysis include high activity, high selectivity, wide substrate scope, and mild reaction conditions. The advantages of electrocatalysis include the possible utilization of renewable electricity as an electron source and high energy conversion efficiency. These properties are integrated to achieve selective biosensing, efficient energy conversion, and the production of diverse products. This review seeks to systematically and comprehensively detail the fundamentals, analyze the existing problems, summarize the development status and applications, and look toward the future development directions of bioelectrocatalysis. First, the structure, function, and modification of bioelectrocatalysts are discussed. Second, the essentials of bioelectrocatalytic systems, including electron transfer mechanisms, electrode materials, and reaction medium, are described. Third, the application of bioelectrocatalysis in the fields of biosensors, fuel cells, solar cells, catalytic mechanism studies, and bioelectrosyntheses of high-value chemicals are systematically summarized. Finally, future developments and a perspective on bioelectrocatalysis are suggested.

Biomedicines

Numerous studies have addressed the utilization of glutaraldehyde (GA) as a homobifunctional cross-linker. However, its applicability has been impeded due to several issues, including the tendency of GA molecules to undergo polymerization. Herein, a portable urea biosensor was developed for the real-time monitoring of the flow of physiological fluids; this was achieved by using disuccinimidyl cross-linker-based urease immobilization. Urease was immobilized on a porous polytetrafluoroethylene (PTFE) solid support using different disuccinimidyl cross-linkers, namely disuccinimidyl glutarate (DSG), disuccinimidyl suberate (DSS) and bis-N-succinimidyl-(pentaethylene glycol) ester (BS(PEG)5). A urease activity test revealed that DSS exhibited the highest urease immobilizing efficiency, whereas FT-IR analysis confirmed that urease was immobilized on the PTFE membrane via DSS cross-linking. The membrane was inserted in a polydimethylsiloxane (PDMS) fluidic chamber that generated an electro...

Nanotechnology Reviews

Global overpopulation, industrial expansion, and urbanization have generated massive amounts of wastes. This is considered as a significant worldwide challenge that requires an urgent solution. Additionally, remarkable advances in the field of biomedicine have impacted the entire spectrum of healthcare and medicine. This has paved the way for further refining of the outcomes of biomedical strategies toward early detection and treatment of different diseases. Various nanomaterials (NMs) have been dedicated to different biomedical applications including drug delivery, vaccinations, imaging modalities, and biosensors. However, toxicity is still the main factor restricting their use. NMs recycled from different types of wastes present a pioneering approach to not only avoid hazardous effects on the environment, but to also implement circular economy practices, which are crucial to attain sustainable growth. Moreover, recycled NMs have been utilized as a safe, yet revolutionary alternati...

Royal Society Open Science

Gold nanoparticles provide a user-friendly and efficient surface for immobilization of enzymes and proteins. In this paper, we present a novel approach for enzyme bioconjugation to gold nanostars (AuNSs). AuNSs were modified with l -cysteine (Cys) and covalently bound to N -hydroxysulfosuccinimide (sulfo-NHS) activated intermediate glucose oxidase (GOx) to fabricate a stable and sensitive AuNSs–Cys–GOx bioconjugate complex. Such a strategy has the potential for increased attachment affinity without protein adsorption onto the AuNSs surface. Good dispersity in buffer suspension was observed, as well as stability in high ionic environments. Using the AuNSs–Cys–GOx bioconjugates showed greater sensitivity in the measuring of low concentrations of glucose based on plasmonic and colorimetric detection. Such a novel approach for enzyme immobilization can lead to AuNSs–Cys–GOx bioconjugate complexes that can be used as catalytic nanodevices in nanobiosensors based on oxidases in biomedical...

Journal of Electrochemical Science and Technology

Cu-Salen complex was prepared and attached into chitosan (Cs) polymer backbone. Nanocomposite of the synthesized polymer was prepared with functionalized carbon nano-particles (Cs-Cu-sal/C) to modify the electrode surface. The surface morphology of (Cs-Cu-sal/C) nanocomposite film showed a homogeneous distribution of carbon nanoparticles within the polymeric matrix. The cyclic voltammogram of the modified electrode exhibited a redox behavior at-0.1 V vs. Ag/AgCl (3 M KCl) in 0.1 M PB (pH 7) and showed an excellent hydrogen peroxide reduction activity. The Cs-Cu-sal/C electrode displays a linear response from 5×10

Micromachines

Metabolic disorders such as the highly prevalent disease diabetes require constant monitoring. The health status of patients is linked to glucose levels in blood, which are typically measured invasively, but can also be correlated to other body fluids such as sweat. Aiming at a reliable glucose biosensor, an enzymatic sensing layer was fabricated on flexible polystyrene foil, for which a versatile nanoimprinting process for microfluidics was presented. For the sensing layer, a gold electrode was modified with a cysteine layer and glutaraldehyde cross-linker for enzyme conformal immobilization. Chronoamperometric measurements were conducted in PBS buffered glucose solution at two potentials (0.65 V and 0.7 V) and demonstrated a linear range between 0.025 mM to 2mM and an operational range of 0.025 mM to 25 mM. The sensitivity was calculated as 1.76µA/mM/cm2 and the limit of detection (LOD) was calculated as 0.055 mM at 0.7 V. An apparent Michaelis–Menten constant of 3.34 mM (0.7 V) a...

Biotechnology & Biotechnological Equipment

An electrochemical organophosphorus hydrolase-based sensor for direct organophosphorus pesticides determination was developed, analytically characterized and applied for paraoxon quantification. The biosensing platform was constructed by one-step electrodeposition onto the surface of a glassy carbon electrode of a bionanocomposite of chitosan with carbon nanotube (CNTs), hydroxyapatite (HA) and organophosphorus hydrolase. The sensor, optimized with respect to the CNTs and HA load, was characterized by the application of cyclic voltammetry and chronoamperometry. The limit of detection (LOD) was found to be as low as 0.1 mmol L ¡1 , the linear concentration range was extended up to 80 mmol L ¡1 , and the sensitivity was as high as 5.10 nA L mmol ¡1. The sensor possesses enhanced sensitivity towards organophosphorus pesticides (OPs), because of the CNTs and the HA nanoparticles synergistic action.

2019

It is extremely challenging to chemically diagnose disorders of the brain. There is hence great interest in designing and optimizing tools for direct detection of chemical biomarkers implicated in neurological disorders to improve diagnosis and treatment. Tools that are capable of monitoring brain chemicals, neurotransmitters in particular, need to be biocompatible, perform with high spatiotemporal resolution, and ensure high selectivity and sensitivity. Recent advances in electrochemical methods are addressing these criteria; the resulting devices demonstrate great promise for in vivo neurotransmitter detection. None of these devices are currently used for diagnostic purposes, however these cutting-edge technologies are promising more sensitive, selective, faster, and less invasive measurements. Via this review we highlight significant technical advances and in vivo studies, performed in the last 5 years, that we believe will facilitate the development of diagnostic tools for brain...

Sensors, 2016

Food allergens are proteins from nuts and tree nuts, fish, shellfish, wheat, soy, eggs or milk which trigger severe adverse reactions in the human body, involving IgE-type antibodies. Sensitive detection of allergens in a large variety of food matrices has become increasingly important considering the emergence of functional foods and new food manufacturing technologies. For example, proteins such as casein from milk or lysozyme and ovalbumin from eggs are sometimes used as fining agents in the wine industry. Nonetheless, allergen detection in processed foods is a challenging endeavor, as allergen proteins are degraded during food processing steps involving heating or fermentation. Detection of food allergens was primarily achieved via Enzyme-Linked Immuno Assay (ELISA) or by chromatographic methods. With the advent of biosensors, electrochemical affinity-based biosensors such as those incorporating antibodies and aptamers as biorecognition elements were also reported in the literature. In this review paper, we highlight the success achieved in the design of electrochemical affinity biosensors based on disposable screen-printed electrodes towards detection of protein allergens. We will discuss the analytical figures of merit for various disposable screen-printed affinity sensors in relation to methodologies employed for immobilization of bioreceptors on transducer surface.

2021

In this study, screen-printed carbon electrodes (SPCE) were modified with electrochemically reduced graphene oxide (ERGO) (SPCE/ERGO), then polythionine (PTH) film was formed on SPCE/ERGO by electropolymerization technique (SPCE/ERGO/PTH). The optimum number of cycles has been determined for the electropolymerization of PTH on SPCE/ERGO. Gold nanoparticles (AuNP) were formed on SPCE/ERGO/PTH electrodes by electrodeposition method and electrochemical characterizations of SPCE/ERGO/PTH/AuNP electrodes were made by cyclic voltammetry (CV). The electrocatalytic effects of ERGO, PTH and AuNP on the BŞEÜ Fen Bilimleri Dergisi 8(1), 8-23, 2021 BSEU Journal of Science https://doi.org/10.35193/bseufbd.822201 e-ISSN: 2458-7575 (https://dergipark.org.tr/tr/pub/bseufbd) 9 electrooxidation of dopamine (DA), nicotinamide adenine dinucleotide (NADH) and ascorbic acid (AA) were investigated, and PTH was observed to play a role as a mediator for three analytes. The anodic peak potentials of DA, NADH...

Nanomaterials (Basel, Switzerland), 2017

We report a simple, versatile, and rapid method for the fabrication of optically-transparent large-area carbon nanotube (CNT) films via flotation assembly. After solvent-induced assembly, floating films were transferred to a flat supporting substrate to form conductive and transparent CNT film electrodes. The resulting electrodes, with uniform 40 ± 20 nm multi-walled CNT (MWCNT) layers, were characterized by electrochemical and microscopy methods. The flotation method does not require specialized thin-film instrumentation and avoids the need for surfactants and pre-oxidized CNTs which can hamper electrochemical performance. A proof-of-concept nanostructured bioelectrode demonstrating high sensitivity for glucose was developed with an electropolymerized poly(pyrene-adamantane) layer for host-guest immobilization of active β-cyclodextrin tagged GOx enzymes. The polymer provides pyrene groups for cross-linking to CNTs and pendant adamantane groups for binding the β-cyclodextrin groups ...

Chemosensors

Personal biosensors and bioelectronics have been demonstrated for use in out-of-clinic biomedical devices. Such modern devices have the potential to transform traditional clinical analysis into a new approach, allowing patients or users to screen their own health or warning of diseases. Researchers aim to explore the opportunities of easy-to-wear and easy-to-carry sensors that would empower users to detect biomarkers, electrolytes, or pathogens at home in a rapid and easy way. This mobility would open the door for early diagnosis and personalized healthcare management to a wide audience. In this review, we focus on the recent progress made in modern electrochemical sensors, which holds promising potential to support point-of-care technologies. Key original research articles covered in this review are mainly experimental reports published from 2018 to 2020. Strategies for the detection of metabolites, ions, and viruses are updated in this article. The relevant challenges and opportun...

Chemosensors, 2020

A sensitive electrochemical detection method for Salmonella spp. was described, based on the use of Hechtia argentea lectin immobilised on a screen-printed gold electrode. The lectin was extracted from Hechtia argentea, a plant belonging to the Bromeliaceae family. The lectin with molecular weight near 27.4 kDa showed selectivity towards D-mannose, contained on the lipopolysaccharide cell wall of Salmonella spp. Carbohydrate selectivity of the lectin was measured as a change in impedance with respect to concentration. The binding of the bacteria to the biosensor surface increased impedance with increasing concentrations of Salmonella spp., achieving a linear range of detection of 15–2.57 × 107 CFU mL−1, with a limit of detection of 5 CFU mL−1. Increases in impedance were measured using electrochemical impedance spectroscopy and analysed using Nyquist plots. The biosensor was applied in analysis of hen egg samples, and the results were consistent with those obtained using the officia...

Journal of Materials Science, 2017

In this work, we report the production of a layer-by-layer (LbL) film of gold nanoparticles stabilized with carrageenan (carr-AuNPs) interspersed with a conductive polyaniline (PANI) layer. Conventionally, PANI has poor electroactivity in physiological buffers, limiting its using in electrochemical biosensors. The films were prepared on low cost and easy to manufacture flexible gold electrodes (FEAu). Two adsorption sequences were tested for production of the films-PANI/carr-AuNP and carr-AuNP/PANI. The gold nanoparticle size and colloidal stability were characterized. The films were characterized by cyclic voltammetry, UV-visible spectroscopy and atomic force microscopy. The results showed the synergistic effects of the carr-AuNPs (120 nm) and PANI, which improved both the electrochemical response and the stability of the conductive polymer in physiological medium by three times. The presence of the carr-AuNPs in the film caused a significant increase in roughness of the FEAu-modified electrode compared to that of an unmodified electrode, resulting in an increased active electrode area. Studies of film growth by UV-Vis spectroscopy indicated that the deposition mechanisms of both films involved an auto-regulating adsorption process, with the same amount of material adsorbed in each coating step. The PANI/carr-AUNP film showed considerable improvement in stability and conductivity compared to PANI-only films in the physiological environment, which confers advantages for use as a biosensor.

Biosensors

The continued focus on improving the quality of human life has encouraged the development of increasingly efficient, durable, and cost-effective products in healthcare. Over the last decade, there has been substantial development in the field of technical and interactive textiles that combine expertise in electronics, biology, chemistry, and physics. Most recently, the creation of textile biosensors capable of quantifying biometric data in biological fluids is being studied, to detect a specific disease or the physical condition of an individual. The ultimate goal is to provide access to medical diagnosis anytime and anywhere. Presently, alcohol is considered the most commonly used addictive substance worldwide, being one of the main causes of death in road accidents. Thus, it is important to think of solutions capable of minimizing this public health problem. Alcohol biosensors constitute an excellent tool to aid at improving road safety. Hence, this review explores concepts about ...

Materials Research Innovations, 2020

An electrochemical gold nanoparticle sensor was developed for the detection of zinc, cadmium and lead ions, due to its electronic properties provided by the large surface/volume ratio and its easy surface modification. Linear ranges for Zn2+, Cd2+ and Pb2+ solutions were 0.469-3.051 µM, 3.968-68.970 µM and 7.937-137.900 µM, respectively. Sensitivity values were superior for electrode modified with chitosan, melamine and gold nanoparticles, and its microamperes/micromoles ratio were 5.7887 [µA/(µM)], 0.8915 [µA/(µM)] and 0.2221 [µA/(µM)], for Zn2+, Cd2+ and Pb2+ solutions, respectively. Synthetic sample solutions with concentrations higher than 1.481 µM Pb2+/L, 37.04 µM Cd2+/L and 74.07 µM Zn2+/L were analysed with high selectivity for the three ions in the mixed water samples. The results showed relevant enhancement of the electrochemical response signals for the ion-selective membrane sensor when the AuNPs added to the composite, a suitable resolution was reached leading to higher selectivity and better accuracy.

Angewandte Chemie (International ed. in English), 2015

We describe the formation of charge-transfer complexes that feature electron-donating carbon nanodots (CND) and electron-accepting perylenediimides (PDI). The functionalities of PDIs have been selected to complement those of CNDs in terms of electrostatic and π-stacking interactions based on oppositely charged ionic head groups and extended π-systems, respectively. Importantly, the contributions from electrostatic interactions were confirmed in reference experiments, in which stronger interactions were found for PDIs that feature positively rather than negatively charged head groups. The electronic interactions between the components in the ground and excited state were characterized in complementary absorption and fluorescence titration assays that suggest charge-transfer interactions in both states with binding constants on the order of 8×10(4) M(-1) (25 L g(-1) ). Selective excitation of the two components in ultrafast pump probe experiments gave a 210 ps lived charge-separated ...

RSC Advances, 2016

Advantages, drawbacks and trends in nanomaterials for enzyme immobilization.

Chemosensors

The world of sensors is diverse and is advancing at a rapid pace due to the fact of its high demand and constant technological improvements. Electrochemical sensors provide a low-cost and convenient solution for the detection of variable analytes and are widely utilized in agriculture, food, and oil industries as well as in environmental and biomedical applications. The popularity of electrochemical sensing stems from two main advantages: the variability of the reporting signals, such as the voltage, current, overall power output, or electrochemical impedance, and the low theoretical detection limits that originate from the differences in the Faradaic and nonFaradaic currents. This review article attempts to cover the latest advances and applications of electrochemical sensors in different industries. The role of nanomaterials in electrochemical sensor research and advancements is also examined. We believe the information presented here will encourage further efforts on the understa...

Biosensors

Two-dimensional transition metal dichalcogenides (2D TMDs) have gained considerable attention due to their distinctive properties and broad range of possible applications. One of the most widely studied transition metal dichalcogenides is molybdenum disulfide (MoS2). The 2D MoS2 nanosheets have unique and complementary properties to those of graphene, rendering them ideal electrode materials that could potentially lead to significant benefits in many electrochemical applications. These properties include tunable bandgaps, large surface areas, relatively high electron mobilities, and good optical and catalytic characteristics. Although the use of 2D MoS2 nanosheets offers several advantages and excellent properties, surface functionalization of 2D MoS2 is a potential route for further enhancing their properties and adding extra functionalities to the surface of the fabricated sensor. The functionalization of the material with various metal and metal oxide nanostructures has a signifi...

Nanomaterials, 2021

Novel nanoarchitectures based on lignin nanoparticles (LNPs) were designed and realized for electrochemical eco-friendly biosensing development. Two types of lignin nanoparticles were utilized for the modification of a gold bare electrode, namely organosolv (OLNPs) and kraft lignin (KLNPs) nanoparticles, synthetized from a sulfur-free and a sulfur lignin, respectively. The electrochemical behavior of LNP-modified electrodes was studied using two electrochemical techniques, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Compared to the gold bare electrode, an evident decrease in the faradaic current and increase of the ΔEp were observed in cyclic voltammograms. In addition, larger semicircles were registered in Nyquist plots. These results suggest a strong inhibition effect of the electron transfer reaction by LNPs layer, especially in the case of KLNPs. The modified electrodes, properly assembled with concanavalin A (ConA) and glucose oxidase (GOx), were s...

Sensors, 2014

The last decade has witnessed an intensive research effort in the field of electrochemical sensors, with a particular focus on the design of amperometric biosensors for diverse analytical applications. In this context, nanomaterial integration in the construction of amperometric biosensors may constitute one of the most exciting approaches. The attractive properties of nanomaterials have paved the way for the design of a wide variety of biosensors based on various electrochemical detection methods to enhance the analytical characteristics. However, most of these nanostructured materials are not explored in the design of amperometric biosensors. This review aims to provide insight into the diverse properties of nanomaterials that can be possibly explored in the construction of amperometric biosensors.

Analytical and Bioanalytical Chemistry, 2017

To immobilize enzymes at the surface of a nanoparticle-based electrochemical sensor is a common method to construct biosensors for non-electroactive analytes. Studying the interactions between the enzymes and nanoparticle support is of great importance in optimizing the conditions for biosensor design. This can be achieved by using a combination of analytical methods to carefully characterize the enzyme nanoparticle coating at the sensor surface while studying the optimal conditions for enzyme immobilization. From this analytical approach, it was found that controlling the enzyme coverage to a monolayer was a key factor to significantly improve the temporal resolution of biosensors. However, these characterization methods involve both tedious methodologies and working with toxic cyanide solutions. Here we introduce a new analytical method that allows direct quantification of the number of immobilized enzymes (glucose oxidase) at the surface of a gold nanoparticle coated glassy carbon electrode. This was achieved by exploiting an electrochemical stripping method for the direct quantification of the density and size of gold nanoparticles coating the electrode surface and combining this information with quantification of fluorophore-labeled enzymes bound to the sensor surface after stripping off their nanoparticle support. This method is both significantly much faster compared to previously reported methods and with the advantage that this method presented is non-toxic.

Catalysts, 2018

Neuronal activity and brain glucose metabolism are tightly coupled, where triggered neurotransmission leads to a higher demand for glucose. To better understand the regulation of neuronal activity and its relation to high-speed metabolism, development of analytical tools that can temporally resolve the transients of vesicular neurotransmitter release and fluctuations of metabolites such as glucose in the local vicinity of the activated neurons is needed. Here we present an amperometric biosensor design for rapid co-detection of glucose and the neurotransmitter dopamine. The sensor is based on the immobilization of an ultra-thin layer of glucose oxidase on to a gold-nanoparticle-covered carbon fiber microelectrode. Our electrode, by altering the potential applied at the sensor surface, allows for the high-speed recording of both glucose and dopamine. We demonstrate that, even though glucose is electrochemically detected indirectly through the enzymatic product and the electroactive dopamine is sensed directly, when exposing the sensor surface to a mixture of the two analytes, fluctuations in glucose and dopamine concentrations can be visualized with similar speed and at a millisecond time scale. Hence, by minimizing the enzyme coating thickness at the sensor surface, dual detection of glucose and dopamine can be realized at the same sensor surface and at time scales necessary for monitoring fast metabolic alterations during neurotransmission.

International Journal of Molecular Sciences, 2019

The ability of commercial monolayer graphene oxide (GO) and graphene oxide nanocolloids (GOC) to interact with different unicellular systems and biomolecules was studied by analyzing the response of human alveolar carcinoma epithelial cells, the yeast Saccharomyces cerevisiae and the bacteria Vibrio fischeri to the presence of different nanoparticle concentrations, and by studying the binding affinity of different microbial enzymes, like the α-l-rhamnosidase enzyme RhaB1 from the bacteria Lactobacillus plantarum and the AbG β-d-glucosidase from Agrobacterium sp. (strain ATCC 21400). An analysis of cytotoxicity on human epithelial cell line A549, S. cerevisiae (colony forming units, ROS induction, genotoxicity) and V. fischeri (luminescence inhibition) cells determined the potential of both nanoparticle types to damage the selected unicellular systems. Also, the protein binding affinity of the graphene derivatives at different oxidation levels was analyzed. The reported results highl...

Journal of Cellular Biotechnology, 2016

In the era of increasing usage of materials, there is a growing concern of environmental aspects. This has motivated many scientists to research on greener materials and technologies. In this line, several biodegradable materials and biomolecules-based technologies have been developed in the recent years. This has triggered vivacious participation of various biomolecules such as DNA, proteins, enzymes, and cells in the field of 'materials'. Among the various biomolecules, enzymes have gained a significant position due to their applications in numerous fields including nanoparticles synthesis, polymer degradation, enzymatic lithography, biosensing and bio-fuel cells. This mini-review accounts on recent trends in selective fields such as nanoparticles synthesis, enzymatic polymer degradation and enzymatic lithography.

Chemical Society Reviews, 2021

Attomolar analyte sensing in clinical diagnosis, food safety and environmental monitoring achieved by the technological advances in transducers design, bioreceptors functionalization and nanomaterials synthesis.

Biosensors and Bioelectronics, 2020

For in-situ disease markers detection, point-of-care (POC) diagnosis has great advantages in speed and cost compared with traditional techniques. The rapid diagnosis, prognosis, and surveillance of diseases can significantly reduce disease-related mortality and trauma. Therefore, increasing attention has been paid to the POC diagnosis devices due to their excellent diagnosis speed and portability. Over the past ten years, paperbased microfluidic aptasensors have emerged as a class of critical POC diagnosis devices and various aptasensors have been proposed to detect various disease markers. However, most aptasensors need further improvement before they can actually enter the market and be widely used. There is thus an urgent need to sort out the key points of preparing the aptasensors and the direction that needs to be invested in. This review summarizes the representative articles in the development of paper-based microfluidic aptasensors. These works can be divided into paper-based optical aptasensors and paper-based electrochemical aptasensors according to their output signals. Significant focus is applied to these works according to the following three parts: (1) The ingenious design of device structure; (2) Application and synthesis of nanomaterial; (3) The detection principle of the proposed aptasensor. This is a detailed and comprehensive review of paper-based microfluidic aptasensors. The accomplishments and shortcomings of the current aptasensors are outlined, the development direction and the future prospective are given. It is hoped that the research in this review can provide a reference for further development of more advanced, more effective paper-based microfluidic aptasensors for POC disease markers diagnosis.

Biosensors

Electrochemistry is a genuinely interdisciplinary science that may be used in various physical, chemical, and biological domains. Moreover, using biosensors to quantify biological or biochemical processes is critical in medical, biological, and biotechnological applications. Nowadays, there are several electrochemical biosensors for various healthcare applications, such as for the determination of glucose, lactate, catecholamines, nucleic acid, uric acid, and so on. Enzyme-based analytical techniques rely on detecting the co-substrate or, more precisely, the products of a catalyzed reaction. The glucose oxidase enzyme is generally used in enzyme-based biosensors to measure glucose in tears, blood, etc. Moreover, among all nanomaterials, carbon-based nanomaterials have generally been utilized thanks to the unique properties of carbon. The sensitivity can be up to pM levels using enzyme-based nanobiosensor, and these sensors are very selective, as all enzymes are specific for their su...

International Journal of Nanomedicine, 2021

Heavy metals cause considerable environmental pollution due to their extent and non-degradability in the environment. Analysis and trace levels of arsenic, lead, mercury, and cadmium as the most toxic heavy metals show that they can cause various hazards in humans' health. To achieve rapid, high-sensitivity methods for analyzing ultra-trace amounts of heavy metals in different environmental and biological samples, novel biosensors have been designed with the participation of strategies applied in nanotechnology. This review attempted to investigate the novel, sensitive, efficient, cost-benefit, point of care, and userfriendly biosensors designed to detect these heavy metals based on functional mechanisms. The study's search strategies included examining the primary databases from 2015 onwards and various keywords focusing on heavy metal biosensors' performance and toxicity mechanisms. The use of aptamers and whole cells as two important bio-functional nanomaterials is remarkable in heavy metal diagnostic biosensors' bioreceptor design. The application of hybridized nanomaterials containing a specific physicochemical function in the presence of a suitable transducer can improve the sensing performance to achieve an integrated detection system. Our study showed that in addition to both labeled and labelfree detection strategies, a wide range of nanoparticles and nanocomposites were used to modify the biosensor surface platform in the detection of heavy metals. The detection limit and linear dynamic range as an essential characteristic of superior biosensors for the primary toxic metals are studied. Furthermore, the perspectives and challenges facing the design of heavy metal biosensors are outlined. The development of novel biosensors and the application of nanotechnology, especially in real samples, face challenges such as the capability to simultaneously detect multiple heavy metals, the interference process in complex matrices, the efficiency and stability of nanomaterials implemented in various laboratory conditions.

Sensors, 2018

Development of new, faster methods of biosensor construction is a huge challenge for current science and industry. In this work, biosensor construction was carried out using a new soft plasma polymerization (SPP) method in which a bio-recognition layer of laccase enzyme was polymerized and bonded to a glassy carbon electrode (GCE) substrate under atmospheric pressure with a corona discharge jet. Laccase belongs to the oxidoreductase enzyme group with four copper atoms in its active center. Application of the corona SPP plasma method allows reduction of the time needed for biosensor construction from several hours to minutes. The presented work includes optimization of the laccase bio-recognition layer deposition time, structural studies of the deposited laccase layer, as well as study of the fabricated biosensor applicability for the determination of Rutin in real pharmaceutical samples. This method produces a biosensor with two linear ranges from 0.3 μmol/dm3 to 0.5 μmol/dm3 and fr...

Sensors, 2018

Electrochemical enzyme-linked immunosorbent assay (ELISA)-based immunoassays for cancer biomarker detection have recently attracted much interest owing to their higher sensitivity, amplification of signal, ease of handling, potential for automation and combination with miniaturized analytical systems, low cost and comparative simplicity for mass production. Their developments have considerably improved the sensitivity required for detection of low concentrations of cancer biomarkers present in bodily fluids in the early stages of the disease. Recently, various attempts have been made in their development and several methods and processes have been described for their development, amplification strategies and testing. The present review mainly focuses on the development of ELISA-based electrochemical immunosensors that may be utilized for cancer diagnosis, prognosis and therapy monitoring. Various fabrication methods and signal enhancement strategies utilized during the last few years for the development of ELISA-based electrochemical immunosensors are described.

Nanomaterials (Basel, Switzerland), 2017

This article provides an overview of the recent development of ZnO nanowires (NWs) for chemoresistive sensing. Working mechanisms of chemoresistive sensors are unified for gas, ultraviolet (UV) and bio sensor types: single nanowire and nanowire junction sensors are described, giving the overview for a simple sensor manufacture by multiple nanowire junctions. ZnO NW surface functionalization is discussed, and how this effects the sensing is explained. Further, novel approaches for sensing, using ZnO NW functionalization with other materials such as metal nanoparticles or heterojunctions, are explained, and limiting factors and possible improvements are discussed. The review concludes with the insights and recommendations for the future improvement of the ZnO NW chemoresistive sensing.

ChemElectroChem, 2019

Gold nanostars with an average tip‐to‐tip length of 52±6 nm were functionalized with different capping agents and used as electrode modification materials for protein electrochemistry. Direct electron transfer between cytochrome c and nanostar‐coated pyrolytic graphite electrodes was observed with the protein in solution. The electrochemical response was improved at nanostars functionalized with a 1 : 1 mixture of 11‐mercaptoundecanoic acid and 4‐mercaptobenzoic acid in comparison with gold nanospheres coated with a similar functionalization. Further immobilization of cytochrome c on pyrolytic graphite while conjugated with the same nanostars guaranteed the maintenance of the protein's native properties, whereas direct adsorption on the bare or nanostar‐modified electrodes resulted in an altered conformational state. The pseudo‐peroxidase activity of the altered cytochrome c was enhanced in the presence of the nanostars.

Materials, 2014

Nanotechnology has played a crucial role in the development of biosensors over the past decade. The development, testing, optimization, and validation of new biosensors has become a highly interdisciplinary effort involving experts in chemistry, biology, physics, engineering, and medicine. The sensitivity, the specificity and the reproducibility of biosensors have improved tremendously as a result of incorporating nanomaterials in their design. In general, nanomaterials-based electrochemical immunosensors amplify the sensitivity by facilitating greater loading of the larger sensing surface with biorecognition molecules as well as improving the electrochemical properties of the transducer. The most common types of nanomaterials and their properties will be described. In addition, the utilization of nanomaterials in immunosensors for biomarker detection will be discussed since these biosensors have enormous potential for a myriad of clinical uses. Electrochemical immunosensors provide a specific and simple analytical alternative as evidenced by their brief analysis times, inexpensive instrumentation, lower assay cost as well as good portability and amenability to miniaturization. The role nanomaterials play in biosensors, their ability to improve detection capabilities in low concentration analytes yielding clinically useful data and their impact on other biosensor performance properties will be discussed. Finally, the most common types of electroanalytical detection methods will be briefly touched upon.

Scientific Reports, 2021

In the present article, we developed a highly sensitive label-free electrochemical immunosensor based on NiFe-layered double hydroxides (LDH)/reduced graphene oxide (rGO)/gold nanoparticles modified glassy carbon electrode for the determination of receptor tyrosine kinase-like orphan receptor (ROR)-1. In this electrochemical immunoassay platform, NiFe-LDH/rGO was used due to great electron mobility, high specific surface area and flexible structures, while Au nanoparticles were prepared and coated on the modified electrodes to improve the detection sensitivity and ROR1 antibody immobilizing (ROR1Ab). The modification procedure was approved by using cyclic voltammetry and differential pulse voltammetry based on the response of peak current to the step by step modifications. Under optimum conditions, the experimental results showed that the immunosensor revealed a sensitive response to ROR1 in the range of 0.01-1 pg mL −1 , and with a lower limit of quantification of 10 attogram/mL (10 ag mL −1). Furthermore, the designed immunosensor was applied for the analysis of ROR1 in several serum samples of chronic lymphocytic leukemia suffering patients with acceptable results, and it also exhibited good selectivity, reproducibility and stability. The receptor tyrosine kinase-like orphan receptor 1 (ROR1), an oncofetal antigen, is a Wnt5a receptor and is widely expressed during embryogenesis, whereas it is absent in most adult tissues 1,2. Studies have reported that ROR1 is expressed at a high level in human leukemia and several solid malignancies 3-5 , suggesting that detection of ROR1 as a prognostic biomarker is important for the on time clinical analysis and the long-term treatment monitoring. Up to now, numerous conventional techniques have been used for the evaluation of ROR1, including

Biosensors, 2016

Cytochromes P450 (CYP, P450) are a large family of heme-active-site proteins involved in many catalytic processes, including steroidogenesis. In humans, four primary enzymes are involved in the metabolism of almost all xenobiotics. Among these enzymes, CYP3A4 is responsible for the inactivation of the majority of used drugs which makes this enzyme an interesting target for many fields of research, especially pharmaceutical research. Since the late 1970s, attempts have been made to construct and develop electrochemical sensors for the determination of substrates. This paper is concerned with the establishment of such a CYP3A4-containing biosensor. The sensor was constructed by adsorption of alternating layers of sub-nanometer gold particle-modified PAMAM (poly-amido-amine) dendrimers of generation 4.0, along with the enzyme by a layer-by-layer assembly technique. Atomic force microscopy (AFM), quartz crystal microbalance (QCM), and Fourier-transformed infrared spectroscopy (FTIR) wer...

Biotechnology and Applied Biochemistry, 2020

Echinococcosis is endemic in Mediterranean regions and is found primarily in the liver. Biliary fistula is a common complication, but major biliary duct involvement is very rare, and occurs in 0.47% of patients with hepatic hydatid disease. Cyst rupture causing secondary peritoneal hydatidosis is a rare but serious complication. We report the case of a 27-year-old man with multiple peritoneal and hepatic hydatid cysts. The patient came to our attention with cholestatic jaundice. Imaging exams showed numerous peritoneal cysts and massive hydatid disease of the liver, which involved the hepatic confluence, with destruction of the right hepatic duct and fistula formation to the left hepatic duct. The patient was treated with pre-operative albendazole therapy and radical surgery, which consisted of resection of all peritoneal cysts and extended right hepatectomy with biliary reconstruction. No recurrence was seen on CT investigations on the 12th month following surgery. Radical surgical approach remains the treatment of choice.

İstanbul Gelişim Üniversitesi Sağlık Bilimleri Dergisi

Aim: The detection of cholesterol concentration in the blood is an important parameter in the diagnosis and follow-up of many diseases, including cardiovascular and hepatobiliary diseases. Nowadays, various methods have been used for the routine analysis of cholesterol, including spectrophotometric, high-performance liquid chromatography (HPLC), and amperometric methods. Since biosensors have advantages such as high sensitivity, fast response, low cost, small size, continuous online detection, and reproducible results, it is aimed to develop a biosensor method for cholesterol determination. Methods: Polymer film immobilization was performed on the gold electrode surface electrochemically in an acetonitrile-water medium containing o-phenylenediamine and benzoquinone. Cholesterol oxidase (ChOx, E.C. 1.1.3.6) enzyme was immobilized on the polymer film-immobilized electrode surface by the self-forming monolayer method. Optimization and characterization studies were carried out for the d...

Micro

Our recent development of a wireless humidity sensor system embedded in incontinence products enables new sensor applications to diagnose and supervise geriatric diseases (i.e., age-related diabetes mellitus type II). The measurement of glucose in urine, so-called glucosuria, is an early indicator for an incipient diabetes mellitus disease, whose symptoms are often age-related but misjudged. In this paper, an incontinence glucose sensor is printed with biocompatible ink and Prussian blue as an electron mediator on foil and functionalized with immobilized glucose oxidase. Inkjet printing of multiple layers of Nafion prevents large interference substances from diffusing into the measuring electrode and allows precise adjustment of the linear working range, which is significantly different from blood glucose measurement. Performance tests show the potential to detect minimum glucose values and store the sensor over a prolonged period at room temperature. The printed glucose sensor can ...

ChemInform, 2014

Journal of The Electrochemical Society, 2019

Reactive oxygen species (ROS) exhibit different spatial and temporal distributions as well as concentrations in-and outside the cell, thereby functioning as signaling or pathogen-destroying molecules. Especially the ROS H 2 O 2 is important for the patho/physiological status of an organism. Electrochemistry (EM) and electron spin resonance (ESR)-based techniques allow quantification of H 2 O 2 in artificial and living systems, coping a concentration range from low nM up to mM. Working electrodes for EM are optimized by diverse modifications and, additionally, redox mediators are used. Ultramicroelectrodes allow scanning of single cells to spatially resolve and quantify extracellular H 2 O 2 in real-time. With ESR spectroscopy, • O 2 ¯, but not H 2 O 2 , can be directly determined by spin probes in-and outside of cells in suspensions. Monitoring H 2 O 2 requires formation of intermediate radicals, detectable with spin probes. Low μM [H 2 O 2 ] can thus be assessed specifically. Using suitable spin traps, in-vivo ESR and immuno-spin trapping can visualize different radicals at their respective production sites in small animals, organs and tissues. Here, the redox reaction cascades may interfere with cell metabolism. Optimization of all methods established for H 2 O 2 determination would be favorable to finally combine them for mutual validation. Thus, a deeper insight into cellular ROS metabolism can be obtained.