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Immobilization of biomaterials is a very important task in the development of biofuel cells and biosensors. Some semiconducting metal-oxide-based supporting materials can be used in these bioelectronics-based devices. In this article, we are reviewing some functionalization methods that are applied for the immobilization of biomaterials. The most significant attention is paid to the immobilization of biomolecules on the surface of semiconducting metal oxides. The improvement of biomaterials immobilization on metal oxides and analytical performance of biosensors by coatings based on conducting polymers, self-assembled monolayers and lipid membranes is discussed. [ABSTRACT FROM AUTHOR]

Polymers represent materials that are applied in almost all areas of modern life, therefore, the characterization of polymer layers using different methods is of great importance. In this review, the main attention is dedicated to the non-invasive and label-free optical and acoustic methods, namely spectroscopic ellipsometry (SE) and quartz crystal microbalance with dissipation (QCM-D). The specific advantages of these techniques applied for in situ monitoring of polymer layer formation and characterization, biomolecule immobilization, and registration of specific interactions were summarized and discussed. In addition, the exceptional benefits and future perspectives of combined spectroscopic ellipsometry and QCM-D (SE/QCM-D) in one measurement are overviewed. Recent advances in the discussed area allow us to conclude that especially significant breakthroughs are foreseen in the complementary application of both QCM-D and SE techniques for the investigation of polymer structure and assessment of the interaction between biomolecules such as antigens and antibodies, receptors and ligands, and complementary DNA strands.

Monitoring and tracking infection is required in order to reduce the spread of the coronavirus disease 2019 (COVID-19), induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To achieve this goal, the development and deployment of quick, accurate, and sensitive diagnostic methods are necessary. The determination of the SARS-CoV-2 virus is performed by biosensing devices, which vary according to detection methods and the biomarkers which are inducing/providing an analytical signal. RNA hybridisation, antigen-antibody affinity interaction, and a variety of other biological reactions are commonly used to generate analytical signals that can be precisely detected using electrochemical, electrochemiluminescence, optical, and other methodologies and transducers. Electrochemical biosensors, in particular, correspond to the current trend of bioanalytical process acceleration and simplification. Immunosensors are based on the determination of antigen-antibody interaction, which on some occasions can be determined in a label-free mode with sufficient sensitivity.

Recent challenges in biomedical diagnostics show that the development of rapid affinity sensors is very important issue. Therefore, in this review we are aiming to outline the most important directions of affinity sensors where polymer-based semiconducting materials are applied. Progress in formation and development of such materials is overviewed and discussed. Some applicability aspects of conducting polymers in the design of affinity sensors are presented. The main attention is focused on bioanalytical application of conducting polymers such as polypyrrole, polyaniline, polythiophene and poly(3,4-ethylenedioxythiophene) ortho-phenylenediamine. In addition, some other polymers and inorganic materials that are suitable for molecular imprinting technology are also overviewed. Polymerization techniques, which are the most suitable for the development of composite structures suitable for affinity sensors are presented. Analytical signal transduction methods applied in affinity sensors based on polymer-based semiconducting materials are discussed. In this review the most attention is focused on the development and application of molecularly imprinted polymer-based structures, which can replace antibodies, receptors, and many others expensive affinity reagents. The applicability of electrochromic polymers in affinity sensor design is envisaged. Sufficient biocompatibility of some conducting polymers enables to apply them as “stealth coatings” in the future implantable affinity-sensors. Some new perspectives and trends in analytical application of polymer-based semiconducting materials are highlighted.

Charge transfer (CT) is a very important issue in the design of biosensors and biofuel cells. Some nanomaterials can be applied to facilitate the CT in these bioelectronics-based devices. In this review, we overview some CT mechanisms and/or pathways that are the most frequently established between redox enzymes and electrodes. Facilitation of indirect CT by the application of some nanomaterials is frequently applied in electrochemical enzymatic biosensors and biofuel cells. More sophisticated and still rather rarely observed is direct charge transfer (DCT), which is often addressed as direct electron transfer (DET), therefore, DCT/DET is also targeted and discussed in this review. The application of conducting polymers (CPs) for the immobilization of enzymes and facilitation of charge transfer during the design of biosensors and biofuel cells are overviewed. Significant attention is paid to various ways of synthesis and application of conducting polymers such as polyaniline, polypyrrole, polythiophene poly(3,4-ethylenedioxythiophene). Some DCT/DET mechanisms in CP-based sensors and biosensors are discussed, taking into account that not only charge transfer via electrons, but also charge transfer via holes can play a crucial role in the design of bioelectronics-based devices. Biocompatibility aspects of CPs, which provides important advantages essential for implantable bioelectronics, are discussed.

Fast and sensitive determination of biologically active compounds is very important in biomedical diagnostics, the food and beverage industry, and environmental analysis. In this review, the most promising directions in analytical application of conducting polymers (CPs) are outlined. Up to now polyaniline, polypyrrole, polythiophene, and poly(3,4-ethylenedioxythiophene) are the most frequently used CPs in the design of sensors and biosensors; therefore, in this review, main attention is paid to these conducting polymers. The most popular polymerization methods applied for the formation of conducting polymer layers are discussed. The applicability of polypyrrole-based functional layers in the design of electrochemical biosensors and biofuel cells is highlighted. Some signal transduction mechanisms in CP-based sensors and biosensors are discussed. Biocompatibility-related aspects of some conducting polymers are overviewed and some insights into the application of CP-based coatings for the design of implantable sensors and biofuel cells are addressed. New trends and perspectives in the development of sensors based on CPs and their composites with other materials are discussed.

The coronavirus disease 2019 (COVID-19) outbreak caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was proclaimed a global pandemic in March 2020. Reducing the dissemination rate, in particular by tracking the infected people and their contacts, is the main instrument against infection spreading. Therefore, the creation and implementation of fast, reliable and responsive methods suitable for the diagnosis of COVID-19 are required. These needs can be fulfilled using affinity sensors, which differ in applied detection methods and markers that are generating analytical signals. Recently, nucleic acid hybridization, antigen-antibody interaction, and change of reactive oxygen species (ROS) level are mostly used for the generation of analytical signals, which can be accurately measured by electrochemical, optical, surface plasmon resonance, field-effect transistors, and some other methods and transducers. Electrochemical biosensors are the most consistent with the general trend towards, acceleration, and simplification of the bioanalytical process. These biosensors mostly are based on the determination of antigen-antibody interaction and are robust, sensitive, accurate, and sometimes enable label-free detection of an analyte. Along with the specification of biosensors, we also provide a brief overview of generally used testing techniques, and the description of the structure, life cycle and immune host response to SARS-CoV-2, and some deeper details of analytical signal detection principles.

In this research a Hg2+ ion biosensor was developed by combining Prussian blue (PB) with glucose oxidase (GOx) – an enzyme that can be inhibited by Hg2+ ions. An application of PB in the design of Hg2+ ion biosensor enabled detecting changes in hydrogen peroxide reduction current at low operational potential of 0.2 vs Ag|AgCl,KClsat. The described Hg2+ ion biosensor exhibited wide linear range from 27 μM to 247 μM of Hg2+ and higher maximal detectable concentration of Hg2+ than other GOx inhibition‐based biosensors, making it convenient for the analysis of samples with high concentration of Hg2+ ions. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

In this article, we report the development of an electrochemical biosensor for the determination of the SARS-CoV-2 spike protein (rS). A gold disc electrode was electrochemically modified to form the nanocrystalline gold structure on the surface. Then, it was further altered by a self-assembling monolayer based on a mixture of two alkane thiols: 11-mercaptoundecanoic acid (11-MUA) and 6-mercapto-1-hexanol (6-MCOH) (SAMmix). After activating carboxyl groups using a N-(3-dimethylaminopropyl)-N'-ethyl-carbodiimide hydrochloride and N-hydroxysuccinimide mixture, the rS protein was covalently immobilized on the top of the SAMmix. This electrode was used to design an electrochemical sensor suitable for determining antibodies against the SARS-CoV-2 rS protein (anti-rS). We assessed the association between the immobilized rS protein and the anti-rS antibody present in the blood serum of a SARS-CoV-2 infected person using three electrochemical methods: cyclic voltammetry, differential pulse voltammetry, and potential pulsed amperometry. The results demonstrated that differential pulse voltammetry and potential pulsed amperometry measurements displayed similar sensitivity. In contrast, the measurements performed by cyclic voltammetry suggest that this method is the most sensitive out of the three methods applied in this research. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

In recent years, many efforts have been made to develop rapid, sensitive and user-friendly glucose biosensors for monitoring blood glucose concentration in patients. In this study, the electrochemical glucose biosensors based on graphite rod (GR) electrode electrochemically modified with dendritic gold nanostructures (DGNs) and glucose oxidase (GOx) were developed. Phenazine methosulfate was used as a soluble redox mediator. Three GOx immobilization methods: adsorption on DGNs and cross-linking with glutaraldehyde (GA) vapour (GA-GOx/DGNs/GR), covalent immobilization on DGNs modified with 11-mercaptoundecanoic acid self-assembled monolayer (SAM) (GOx-SAM/DGNs/GR) and covalent immobilization on SAM with additional cross-linking with GA vapour (GA-GOx-SAM/DGNs/GR), were used. It was determined that GA significantly improved the stability of the enzyme layer. The difference of maximal current generated during the enzymatic reaction (ΔImax) equal to 272.06 ± 8.69 µA was obtained using a biosensor based on GA-GOx/DGNs/GR electrodes. However, the highest ΔImax equal to 384.20 ± 16.06 µA was obtained using GA-GOx-SAM/DGNs/GR electrode. ΔImax for biosensors based on the GA-GOx-SAM/DGNs/GR electrode was 1.41 times higher than for the GA-GOx/DGNs/GR, whereas the linear dynamic range from 0.1 to 10 mM was the same using all three GOx immobilization methods. The limit of detection using GA-GOx-SAM/DGNs/GR and GA-GOx/DGNs/GR electrodes was 0.019 and 0.022 mM, respectively. The ability to detect glucose in the serum by developed biosensors was evaluated. [ABSTRACT FROM AUTHOR]

Scanning electrochemical microscopy (SECM) is an emerging electroanalytical sensing technique, used to investigate the electrochemical properties of the sample by ultra‐micro‐electrode(UME) scanning probe. UME signal usually is the current, which depends not only on the properties of the evaluated system but also on UME characteristics such as geometry. Variations of UME geometry can decrease accuracy of the measurement, and then correct analysis of the SECM data becomes almost impossible. In the present work, we studied the precision of measurements with three different the most frequent types of defected UME's ((i) recessed‐UME, (ii) outwarded‐UME, (iii) cone‐UME). Measurement results were compared with that obtained with not defected standard‐plane‐UME. Computational experiment was performed with SECM model using diffusion equations with non‐rectangular border conditions to calculate estimated currents for these three types of defected UMEs and to compare them with that for standard‐plane‐UME. In order to test the correctness of the model, computations for recessed‐UME model were compared with data of real‐recessed‐UME experiment. [ABSTRACT FROM AUTHOR]

In this work, three types of electrodes suitable for amperometric glucose biosensors were designed. One type of electrode was based on bio‐selective layer of polypyrrole/(glucose oxidase)/(Prussian Blue) (Ppy/GOx/PB) and it was used as a control electrode regarding to which electrochemical properties of two other types of electrodes were compared. During the formation of Prussian blue layers graphite electrodes were additionally modified by Ni‐hexacyanoferrate (NiHCF) and by Co‐hexacyanoferrate (CoHCF) in order to design Ppy/GOx/PB‐NiHCF and Ppy/GOx/PB‐CoHCF electrodes, respectively. Some physicochemical characteristics of all three types of electrodes were evaluated and compared. The Ppy/GOx/PB‐NiHCF electrode showed wider linear range of the calibration curve than Ppy/GOx/PB and Ppy/GOx/PB‐CoHCF electrodes. The effect of temperature on analytical performance of the Ppy/GOx/PB‐NiHCF based biosensor has been evaluated and activation energy of enzyme catalysed reaction has been calculated within the temperature range of 15 °C to 30 °C. [ABSTRACT FROM AUTHOR]

Abstract: A newly developed amperometric glucose biosensor based on graphite rod (GR) working electrode modified with biocomposite consisting of poly (pyrrole‐2‐carboxylic acid) (PCPy) particles and enzyme glucose oxidase (GOx) was investigated. The PCPy particles were synthesized by chemical oxidative polymerization technique using H2O2 as initiator of polymerization reaction and modified covalently with the GOx (PCPy‐GOx) after activation of carboxyl groups located on the particles surface with a mixture of N‐(3‐dimethylaminopropyl)‐N′‐ethylcarbodiimide hydrochloride (EDC) and N‐hydroxysuccinimide (NHS). Then the PCPy‐GOx biocomposite was dispersed in a buffer solution containing a certain amount of bovine serum albumin (BSA). The resulting biocomposite suspension was adsorbed the on GR electrode surface with subsequent solvent airing and chemical cross‐linking of the proteins with glutaraldehyde vapour (GR/PCPy‐GOx). It was determined that the current response of the GR/PCPy‐GOx electrodes to glucose measured at +300 mV vs Cl reference electrode was influenced by the duration of the PCPy particles synthesis, pH of the GOx solution used for the PCPy particles modification and the amount of immobilized PCPy‐GOx biocomposite. An optimal pH of buffer solution for operation of the biosensor was found to be 8.0. Detection limit was determined as 0.039 mmol L−1 according signal to noise ratio (S/N: 3). The proposed glucose biosensor was tested in human serum samples. [ABSTRACT FROM AUTHOR]

Achievements in polymer chemistry enables to design artificial phase boundaries modified by imprints of selected molecules and some larger structures. These structures seem very useful for the design of new materials suitable for affinity chromatography and sensors. In this review, we are overviewing the synthesis of molecularly imprinted polymers (MIPs) and the applicability of these MIPs in the design of affinity sensors. Such MIP-based layers or particles can be used as analyte-recognizing parts for sensors and in some cases they can replace very expensive compounds (e.g.: antibodies, receptors etc.), which are recognizing analyte. Many different polymers can be used for the formation of MIPs, but conducing polymers shows the most attractive capabilities for molecular-imprinting by various chemical compounds. Therefore, the application of conducting polymers (e.g.: polypyrrole, polyaniline, polythiophene, poly(3,4-ethylenedioxythiophene), and ortho-phenylenediamine) seems very promising. Polypyrrole is one of the most suitable for the development of MIP-based structures with molecular imprints by analytes of various molecular weights. Overoxiation of polypyrrole enables to increase the selectivity of polypyrrole-based MIPs. Methods used for the synthesis of conducting polymer based MIPs are overviewed. Some methods, which are applied for the transduction of analytical signal, are discussed, and challenges and new trends in MIP-technology are foreseen. [Display omitted] • Principles of molecular imprinting are discussed • Synthesis of molecularly imprinted polymers (MIPs) is overviewed • Special attention is dedicated to application in MIP-technology some overoxidized CPs • Applicability of MIPs in affinity sensor design is discussed • Bio-compatibility of some conducting polymers is outlined [ABSTRACT FROM AUTHOR]

Recent challenges in the pharmaceutical and biomedical fields require the development of new analytical methods. Therefore, the development of new sensors is a very important task. In this paper, we are outlining the development of molecularly imprinted polymer (MIP) based sensors, which belongs to important branch of affinity sensors. In this review, recent advances in the design of MIP-based sensors are overviewed. MIPs-based sensing structures can replace expensive natural affinity compounds such as receptors or antibodies. Among many different polymers, conducting polymers show the most versatile properties, which are suitable for sensor application. Therefore, significant attention is paid towards MIPs based on conducting polymers, namely polypyrrole, polythiophene, poly(3,4-ethylenedioxythiophene), polyaniline and ortho-phenylenediamine. Moreover, many other materials, which could be imprinted analyte molecules, are overviewed. Among many conducting polymers, polypyrrole is highlighted as one of the most suitable for molecular imprinting. Some attention is dedicated to overview polymerization methods applied for the design of sensing structures used in various affinity sensors. The transduction of analytical signal is an important issue, therefore, physicochemical methods suitable for analytical signal transduction are also outlined. Advances, trends and perspectives in MIP application are discussed. [Display omitted] • Formation and application of molecularly imprinted polymers (MIPs) is outlined. • Synthesis of conducting polymers modified by molecular imprints is overviewed. • Significant attention is paid towards imprinting of large molecular weight compound such as proteins. • Application of overoxidation of polypyrrole for formation of advanced molecular imprints is discussed. • Analytical signal transduction systems are overviewed. [ABSTRACT FROM AUTHOR]

Graphite rod (GR) modified with electrochemicaly deposited gold nanoparticles (AuNPs) and adsorbed glucose oxidase (GOx) was used in amperometric glucose biosensor design. Enzymatic formation of polypyrrole (Ppy) on the surface of GOx/AuNPs/GR electrode was applied in order to improve analytical characteristics and stability of developed biosensor. The linear glucose detection range for Ppy/GOx/AuNPs/GR electrode was dependent on the duration of Ppy-layer formation and the linear interval was extended up to 19.9 mmol L−1 after 21 h lasting synthesis of Ppy. The sensitivity of the developed biosensor was determined as 21.7 μA mM−1 cm−2, the limit of detection - 0.20 mmol L−1. Ppy/GOx/AuNPs/GR electrodes demonstrated advanced good stability (the t1/2 was 9.8 days), quick detection of glucose (within 5 s) in the wide linear interval. Additionally, formed Ppy layer decreased the influence of electroactive species on the analytical signal. Developed biosensor is suitable for the determination of glucose in human serum samples. [ABSTRACT FROM AUTHOR]

In this research single-step procedure for the modification of graphite electrode by polypyrrole (PPy), Prussian blue (PB) and glucose oxidase (GOx) based composite layer (PPy/PB/GOx) is reported. The PPy/PB/GOx-modified electrode was applied in the design of glucose biosensor. Optimal potential for the registration of amperometric response towards glucose was at +0.05 V vs Ag/AgCl (KClsat.) . Preliminary optimization of initial concentration of pyrrole in initial solution, which was used for the formation of PPy/PB/GOx layer, was performed. The sensitivity of several different PPy/PB/GOx-modified graphite electrodes, which were modified by PPy/PB/GOx layer at different pyrrole concentrations, was evaluated, and it was in a range of 1.0–1.9 μA cm −2 mM −1 . [ABSTRACT FROM AUTHOR]

In this research, scanning electrochemical microscopy combined with electrochemical impedance spectroscopy has been applied to irreversible electroporation of active yeast cells by causing cell death. This finding is important for the development of irreversible electroporation technique, which could be suitable for the curing of cancerous tissues, because during this research cell death has been achieved using relatively low ultramicro-electrode (UME) voltage, precisely of 2.0 V vs Ag/AgCl,Cl-sat. It was determined that the irreversibly electroporated area of immobilized yeast cells was located directly below the UME and was of approximately 20 times larger width than the diameter of the UME, leaving undamaged cells out of this area. The ability of SECM to move the UME with high accuracy in x, y, and z directions and the ability to use electrodes of various diameters as well as the fact that the diameter of the electroporated area depends on the diameter of the UME and on the distance between the UME and the surface, what offers the possibility to establish targeted electroporation systems for selective treatment of tissues. [Display omitted] [ABSTRACT FROM AUTHOR]

Zinc oxide (ZnO) nanostructures are widely used in optical sensors and biosensors. Functionalization of these nanostructures with polymers enables optical properties of ZnO to be tailored. Polydopamine (PDA) is a highly biocompatible polymer, which can be used as a versatile coating suitable for application in sensor and biosensor design. In this research, we have grown ZnO-based nanorods on the surface of ITO-modified glass-plated optically transparent electrodes (glass/ITO). Then the deposition of the PDA polymer layer on the surface of ZnO nanorods was performed from an aqueous PDA solution in such a way glass/ITO/ZnO-PDA structure was formed. The ZnO-PDA composite was characterized by SEM, TEM, and FTIR spectroscopy. Then glucose oxidase (GOx) was immobilized using crosslinking by glutaraldehyde on the surface of the ZnO-PDA composite, and glass/ITO/ZnO-PDA/GOx-based biosensing structure was designed. This structure was applied for the photo-electrochemical determination of glucose (Glc) in aqueous solutions. Photo-electrochemical determination of glucose by cyclic voltammetry and amperometry has been performed by glass/ITO/ZnO-PDA/GOx-based biosensor. Here reported modification/functionalization of ZnO nanorods with PDA enhances the photo-electrochemical performance of ZnO nanorods, which is well suited for the design of photo-electrochemical sensors and biosensors. [ABSTRACT FROM AUTHOR]

Here, we report a novel enantioselective electrochemical biosensor for the discrimination of cystine enantiomers (D- and L-cystine) using a chiral interface for the specific recognition of D- and L-cystine. The biosensor is based on reduced graphene oxide modified by ß-cyclodextrin (rGO/ß-CD) at the GCE surface. During the preparation of rGO/ß-CD/GCE, the modified electrode surfaces were characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). The electrochemical behaviours of the D- and L-cystine were investigated using the rGO/ß-CD/GCE by CV and compared to bare GCE. A clear separation between the oxidation peak potentials of D- and L-cystine was observed at 1.32 and 1.42 V, respectively. The electrochemical discrimination performance of the fabricated chiral sensor was also examined by differential pulse voltammetry (DPV) in a mixed solution of D- and L-cystine. In addition, the DPV technique was used for the determination of D- and L-cystine at low concentration values in the range of 1.0-10.0 μM. To investigate the amperometric response of rGO/ß-CD/GCE towards D- and L-cystine, the chronoamperometry technique was used in the concentration range of 10.0-100.0 μM. The interactions of the enantiomers with rGO/ß-CD were modelled by molecular docking using AutoDock Vina, and the interaction energies were predicted to be -4.8 and -5.3 kcal mol-1 for D- and L-cystine, respectively. The corresponding values of binding constants were calculated to be 3.32 x 10³ and 7.71 x 10³ M-1, respectively. The experimental and molecular docking results indicate that the rGO/ß-CD/GCE has a different affinity for each enantiomer. [ABSTRACT FROM AUTHOR]

The electrical field-induced changes of the yeast Saccharomyces cerevisiae cells permeabilization to tetraphenylphosphonium (TPP+) ions were studied using square-shaped, nanosecond duration high power electrical pulses. It was obtained that pulses having durations ranging from 10 ns to 60 ns, and generating electric field strengths up to 190 kV/cm significantly (up to 65 times) increase the absorption rate of TPP+ ions without any detectible influence on the yeast cell viability. The modelling of the TPP+ absorption process using a second order rate equation demonstrates that depending on the duration of the pulses, yeast cell clusters of different sizes are homogeniously permeabilized. It was concluded, that nanosecond pulse-induced permeabilization can be applied to increase the operational speed of whole cell biosensors. [ABSTRACT FROM AUTHOR]

A biosensor based on glucose oxidase (GOx) immobilized on gold nanoparticles (Au-NPs) electrochemically predeposited on the surface of graphite rod (GR) electrode was developed (GOx/Au-NPs/GR). Main analytical characteristics of this biosensor were determined and compared with those determined using a biosensor setup without Au-NP modification (GOx/GR). The highest analytical signal of GOx/Au-NPs/GR electrode was observed after 13 nm Au-NP deposition on the electrode from 0.8 nmol L-1 solution lasting 20 min, when cyclic voltammetry was performed in the range from 0.0 to +1.0 V vs Ag/AgCl. The best analytical characteristics of the developed biosensor were obtained after 25 mg mL-1 GOx immobilization on the Au-NPs/GR electrode. Analytical signal registered using GOx/Au-NPs/GR electrode was 2.08 times higher in comparison to GOx/GR electrode. The registered currents of both electrodes were linearly dependent on glucose concentration in the range of 0.1-10 mmol L-1. The developed GOx/Au-NPs/GR electrode was characterized by high sensitivity, which was equal to 101.02 μA mM-1 cm-2 in the linear glucose detection range. The limit of detection was 0.083 mmol L-1 with relative standard deviation of 6% for GOx/Au-NPs/GR electrode. This study demonstrates a successful practical exploitation of the developed biosensor in a human serum sample. [ABSTRACT FROM AUTHOR]

This review focuses on the overview of microbial amperometric biosensors and microbial biofuel cells (MFC) and shows how very similar principles are applied for the design of both types of these bioelectronics-based devices. Most microorganism-based amperometric biosensors show poor specificity, but this drawback can be exploited in the design of microbial biofuel cells because this enables them to consume wider range of chemical fuels. The efficiency of the charge transfer is among the most challenging and critical issues during the development of any kind of biofuel cell. In most cases, particular redox mediators and nanomaterials are applied for the facilitation of charge transfer from applied biomaterials towards biofuel cell electrodes. Some improvements in charge transfer efficiency can be achieved by the application of conducting polymers (CPs), which can be used for the immobilization of enzymes and in some particular cases even for the facilitation of charge transfer. In this review, charge transfer pathways and mechanisms, which are suitable for the design of biosensors and in biofuel cells, are discussed. Modification methods of the cell-wall/membrane by conducting polymers in order to enhance charge transfer efficiency of microorganisms, which can be potentially applied in the design of microbial biofuel cells, are outlined. The biocompatibility-related aspects of conducting polymers with microorganisms are summarized. [ABSTRACT FROM AUTHOR]

Recent challenges in biomedical diagnostics show that the development of rapid affinity sensors is very important issue. Therefore, in this review we are aiming to outline the most important directions of affinity sensors where polymer-based semiconducting materials are applied. Progress in formation and development of such materials is overviewed and discussed. Some applicability aspects of conducting polymers in the design of affinity sensors are presented. The main attention is focused on bioanalytical application of conducting polymers such as polypyrrole, polyaniline, polythiophene and poly(3,4-ethylenedioxythiophene) ortho-phenylenediamine. In addition, some other polymers and inorganic materials that are suitable for molecular imprinting technology are also overviewed. Polymerization techniques, which are the most suitable for the development of composite structures suitable for affinity sensors are presented. Analytical signal transduction methods applied in affinity sensors based on polymer-based semiconducting materials are discussed. In this review the most attention is focused on the development and application of molecularly imprinted polymer-based structures, which can replace antibodies, receptors, and many others expensive affinity reagents. The applicability of electrochromic polymers in affinity sensor design is envisaged. Sufficient biocompatibility of some conducting polymers enables to apply them as "stealth coatings" in the future implantable affinity-sensors. Some new perspectives and trends in analytical application of polymer-based semiconducting materials are highlighted. [ABSTRACT FROM AUTHOR]

Charge transfer (CT) is a very important issue in the design of biosensors and biofuel cells. Some nanomaterials can be applied to facilitate the CT in these bioelectronics-based devices. In this review, we overview some CT mechanisms and/or pathways that are the most frequently established between redox enzymes and electrodes. Facilitation of indirect CT by the application of some nanomaterials is frequently applied in electrochemical enzymatic biosensors and biofuel cells. More sophisticated and still rather rarely observed is direct charge transfer (DCT), which is often addressed as direct electron transfer (DET), therefore, DCT/DET is also targeted and discussed in this review. The application of conducting polymers (CPs) for the immobilization of enzymes and facilitation of charge transfer during the design of biosensors and biofuel cells are overviewed. Significant attention is paid to various ways of synthesis and application of conducting polymers such as polyaniline, polypyrrole, polythiophene poly(3,4-ethylenedioxythiophene). Some DCT/DET mechanisms in CP-based sensors and biosensors are discussed, taking into account that not only charge transfer via electrons, but also charge transfer via holes can play a crucial role in the design of bioelectronics-based devices. Biocompatibility aspects of CPs, which provides important advantages essential for implantable bioelectronics, are discussed. [ABSTRACT FROM AUTHOR]

Abstract: This research represents the evaluation of electrochemical formation of conducting polymer polypyrrole-based composite layer, which could be applied in immunosensor design. Polypyrrole (Ppy) layer was formed by mean of potential pulses and bovine leukaemia virus (BLV) protein gp51 (gp51) was entrapped during this synthesis within formed Ppy layer (gp51/Ppy). Some Ppy layer formation aspects were evaluated and mathematical model, which is describing tendencies of gp51/Ppy layer formation, was adopted. The interaction of gp51/Ppy layer with specific antibodies that are present in the blood serum of BLV infected cattle was evaluated by pulsed amperometric detection. [Copyright &y& Elsevier]

In this study, two ortho-quinoidal compounds, 1,10-phenanthroline-5,6-dione (PD) and 9,10-phenanthrenequinone (PQ), were examined as electron transfer mediators suitable for amperometric glucose biosensors. The dependences of the electrochemical responses of PD- and PQ-based amperometric glucose biosensors on varied concentrations of glucose were investigated under aerobic and anaerobic conditions. The PD-modified graphite rod (GR) electrode revealed a current response seven times higher than that of the PQ-modified GR electrode. The reactivity indices of ortho-quinoidals assessed by means of B3LYP functional method applying 6-311G(D) basis set showed that the electron-accepting potency for PD was markedly higher as compared with that of PQ. Compared to PQ, considerably higher reactivity of PD has been defined in the reactions with NADP-ferredoxin reductase (FNR, EC 1.18.1.2) as a model single-electron transfer FAD-dependent enzyme, which provided an additional evidence for PD as a more efficient mediator compared to PQ. This study illustrates that PD can be applied as a redox mediator for glucose oxidase and it could be more suitable for a reagent-less biosensor design than PQ. [ABSTRACT FROM AUTHOR]

Glucose oxidase (GOx) from Penicillium funiculosum 46.1 was purified using step-by-step ultrafiltration and it was characterized by spectrophotometric and spectrofluorometric methods. It was shown that spectra of GOx produced by P. funiculosum are typical for flavoproteins. Absorption spectrum has distinct peaks at 380 and 457 nm, excitation spectrum at 373 and 447 nm, and emission spectrum at 530 and 562 nm. The pH correlation of enzyme activity and catalytic characteristics in various buffer systems (phosphate (pH 5.0–9.0), citrate (pH 3.0–5.0), citrate-phosphate (pH 3.0–9.0), and universal (pH 3.0–9.0)) were registered. It was determined that the GOx is the most efficiently interacting with substrate (glucose) in phosphate buffer at pH 7.0 with kcat/ Km = 21,825 M −1 s −1. Interaction of several different redox mediators (9,10-phenantroline-5,6-dione, 9,10-phenanthrenequinone, N-methylphenazonium methyl sulfate, ferrocene, ferrocenecarboxylic acid, α-methylferrocenemethanol, ferrocenecarboxaldehyde) with GOx from P. funiculosum was investigated by evaluation of the difference in fluorescence emission intensity of FAD (oxidized) and FADH 2(reduced) forms. It was found that 9,10-phenantroline-5,6-dione and 9,10-phenanthrenequinone are the best redox mediators for this type of GOx. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Fast and sensitive determination of biologically active compounds is very important in biomedical diagnostics, the food and beverage industry, and environmental analysis. In this review, the most promising directions in analytical application of conducting polymers (CPs) are outlined. Up to now polyaniline, polypyrrole, polythiophene, and poly(3,4-ethylenedioxythiophene) are the most frequently used CPs in the design of sensors and biosensors; therefore, in this review, main attention is paid to these conducting polymers. The most popular polymerization methods applied for the formation of conducting polymer layers are discussed. The applicability of polypyrrole-based functional layers in the design of electrochemical biosensors and biofuel cells is highlighted. Some signal transduction mechanisms in CP-based sensors and biosensors are discussed. Biocompatibility-related aspects of some conducting polymers are overviewed and some insights into the application of CP-based coatings for the design of implantable sensors and biofuel cells are addressed. New trends and perspectives in the development of sensors based on CPs and their composites with other materials are discussed. [ABSTRACT FROM AUTHOR]

Polymers represent materials that are applied in almost all areas of modern life, therefore, the characterization of polymer layers using different methods is of great importance. In this review, the main attention is dedicated to the non-invasive and label-free optical and acoustic methods, namely spectroscopic ellipsometry (SE) and quartz crystal microbalance with dissipation (QCM-D). The specific advantages of these techniques applied for in situ monitoring of polymer layer formation and characterization, biomolecule immobilization, and registration of specific interactions were summarized and discussed. In addition, the exceptional benefits and future perspectives of combined spectroscopic ellipsometry and QCM-D (SE/QCM-D) in one measurement are overviewed. Recent advances in the discussed area allow us to conclude that especially significant breakthroughs are foreseen in the complementary application of both QCM-D and SE techniques for the investigation of polymer structure and assessment of the interaction between biomolecules such as antigens and antibodies, receptors and ligands, and complementary DNA strands. [ABSTRACT FROM AUTHOR]

Abstract: Various nanotechnological methods and high number of nanostructured materials including polymer nanocomposites (PNCs) become extremely important in sensor and biosensor technology. Conducting polymers as artificial versatile materials seem very suitable for the development of various analyte-recognizing parts of sensors and biosensors. This contribution is reviewing major fabrication methods of conducting polymer-based nanocomposites for biosensorics. Experience of authors in application of various electrochemically generated polymers in design of biosensors is presented. The influence of fabrication method on sensor characteristics is overviewed. Major types of biosensors based on conducting polymers including catalytic biosensors, DNA-sensors, immunosensors and molecularly imprinted polymer-based affinity sensors is discussed. Conducting and electrochemically generated polymers that are mostly used in sensor design are presented. [Copyright &y& Elsevier]

Abstract: In this study electrochemistry of glucose oxidase (GOx) immobilized on a carbon rod electrode modified by gold nanoparticles (Au-NPs) and π-π conjugated polymer - polypyrrole (Ppy) was studied. Au-NPs facilitated indirect electron transfer via red-ox mediator and showed a positive effect on the amperometric signals of such type electrodes. Several types of electrodes modified by GOx and Au-NPs of different sizes and additionally covered by Ppy layer were investigated. The sensitivity and stability of developed biosensors were evaluated and compared. [Copyright &y& Elsevier]

Abstract: Different glucose biosensors based on glucose oxidase (GOx) immobilised on bare carbon rod electrode (CR) modified with gold nanoparticles (Au-NPs) of (i) 3.5nm (GOx/3.5Au-NPs/CR), (ii) 6nm (GOx/6Au-NPs/CR) and (iii) 13nm (GOx/13Au-NPs/CR) were investigated and compared with biosensors based on GOx immobilised on bare CR (GOx/CR). Enzymatic polymerisation of pyrrole was applied to increase linear detection range of biosensors. The influence of the formed polypyrrole layer on sensitivity and Michaelis–Menten kinetics of designed electrochemical biosensors was investigated. The linear glucose detection interval for GOx/CR and GOx/Au-NPs/CR electrodes was dependent on the duration of polymerisation. [Copyright &y& Elsevier]

The aim of this work was to further evaluate the effect of colloidal solutions of gold nanoparticles (Au-NPs) on the performance of a carbon rod electrode modified with glucose oxidase. The amperometric response of the system at +0.3 V vs. Ag/AgCl was studied in the absence and in the presence of Au-NPs (6 nm and 13 nm in diameter) and in presence of N-methylphenazonium methyl sulfate (PMS) at pH 6.0. This study shows that the application of <0.60 nmol L concentrations of Au-NPs increases the rate of mediated electron transfer, and this effect does not depend on the diameter of Au-NPs. The analytical signal in the presence 0.60 nmol L of Au-NPs (13 nm) and 2 mmol L of PMS linearly depends on the concentration of glucose in the range from 0.1 to 10 mmol L, the limit of detection is as low as 0.05 mmol L. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Abstract: Polypyrrole (Ppy) has been shown as a matrix for label-free electrochemical immunosensor based on electrochemical impedance spectroscopy (EIS) measurements. The immunosensing system model presented here was based on bovine leukemia virus (BLV) protein (gp51) entrapped within electrochemically-synthesized polypyrrole (Ppy/gp51). This Ppy/gp51 layer interacted with antibodies against gp51 (anti-gp51-Ab) that are present in significant concentration in the blood serum of BLV infected cattle. After this interaction protein complex (Ppy/gp51/anti-gp51-Ab) was formed. The horseradish peroxidase (HRP) labeled secondary antibodies (Ab⁎) against anti-gp51-Ab were applied as agents interacting with Ppy/gp51/anti-gp51-Ab and forming the large protein complex (Ppy/gp51/anti-gp51-Ab/Ab⁎). The EIS study was performed for electrodes modified with different Ppy layers described here and an optimal equivalent circuit was adopted for evaluation of EIS spectra, it was a major outcome of this study. [Copyright &y& Elsevier]

An affinity sensor based on a capacitive micromachined ultrasound transducer (cMUT) is reported by this Communication. The cMUT micromembrane arrays modified with adsorbed bovine leukemia virus protein gp51 were applied as a biological recognition part. The cMUT-based sensor is shown to be sensitive to the antibodies against bovine leukemia virus protein gp51 (anti-gp51). Two different concentrations of anti-gp51-containing samples and one blank sample without anti-gp51 were tested. The sensitivity of cMUT-based immunosensor is comparable with the sensitivity of a quartz microbalance-based immunosensor. The cMUT array provides a multi-channel system for the measurement of analytical signal. Moreover, two different characteristics – the resonance frequency shift (Δf) and the shift of the real part of the electromechanical impedance (ΔRe) – could have been evaluated simultaneously. Both analytical signals are informative and can be applied for the estimation of immune complex formation. We found the performance of such a system being potentially superior over some other immunosensing techniques. It is more rapid than electrochemical techniques and provides two different informative parameters. [ABSTRACT FROM AUTHOR]

A direct, label-free detection method of antibodies against the human growth hormone (anti-HGH) using a surface plasmon resonance (SPR) biosensor is reported. The sensing surface of the surface plasmon resonance biosensor chip (SPR-chip) was modified by covalent coupling of the human growth hormone (HGH) to the self-assembled monolayer of 11-mercaptoundecanoic acid (MUA). HGH was immobilized viaprimary amine groups after activation of the MUA carboxyl groups with a mixture of N-hydroxysuccinimide (NHS), and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC). The specific binding of monoclonal anti-HGH antibody on the HGH-modified surface was examined in the concentration range from 0.25 nM to 10 µM. The experimentally observed detection minimum for anti-HGH was 2.47 nM. A single immunoassay cycle could be done within 30 min including the HGH and anti-HGH association, HGH/anti-HGH complex dissociation and surface regeneration steps. The SPR biosensor response for repeatable detections of anti-HGH was highly reproducible and very stable. On the SPR-chip the formed HGH and anti-HGH complex (HGH/anti-HGH) could be gently dissociated and the sensing surface might be regenerated by 50 mM NaOH and 0.5% sodium dodecylsulfate (SDS) solution. Any changes in the original baseline level were detected during the 40 detection–regeneration cycles. This means that damage of the immobilized HGH-based sensitive layer during regeneration was minimal. It was demonstrated that the developed SPR-chip could be stored for at least 21 days before use without considerable loss of sensitivity towards anti-HGH. [ABSTRACT FROM AUTHOR]

Monitoring and tracking infection is required in order to reduce the spread of the coronavirus disease 2019 (COVID-19), induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To achieve this goal, the development and deployment of quick, accurate, and sensitive diagnostic methods are necessary. The determination of the SARS-CoV-2 virus is performed by biosensing devices, which vary according to detection methods and the biomarkers which are inducing/providing an analytical signal. RNA hybridisation, antigen-antibody affinity interaction, and a variety of other biological reactions are commonly used to generate analytical signals that can be precisely detected using electrochemical, electrochemiluminescence, optical, and other methodologies and transducers. Electrochemical biosensors, in particular, correspond to the current trend of bioanalytical process acceleration and simplification. Immunosensors are based on the determination of antigen-antibody interaction, which on some occasions can be determined in a label-free mode with sufficient sensitivity. [ABSTRACT FROM AUTHOR]

Abstract: Enzymatic biofuel cell based on enzyme modified anode and cathode electrodes are both powered by ethanol and operate at ambient temperature is described. The anode of the presented biofuel cell was based on immobilized quino-hemoprotein-alcohol dehydrogenase (QH-ADH), while the cathode on co-immobilized alcohol oxidase (AOx) and microperoxidase (MP-8). Two enzymes AOx and MP-8 acted in the consecutive mode and were applied in the design of the biofuel cell cathode. The ability of QH-ADH to transfer electrons directly towards the carbon-based electrode and the ability of MP-8 to accept electrons directly from the same type of electrodes was exploited in this biofuel cell design. Direct electron transfer (DET) to/from enzymes was the basis for generating an electric potential between the anode and cathode. Application of immobilized enzymes and the harvesting of the same type of fuel at both electrodes (cathode and anode) avoided the compartmentization of enzymatic biofuel cell. The maximal open circuit potential of the biofuel cell was 240mV. [Copyright &y& Elsevier]

This study describes a new, basic procedure for the tuning of some analytical parameters of enzymatic biosensors that are based on hydrogen peroxide-producing oxido-reductases. An amperometric biosensor based on glucose oxidase (GOx) (EC 1.1.3.4) from Penicillum vitale, immobilized on a carbon rod electrode by cross-linking with glutaraldehyde, was exploited as a model system for demonstration of the approach described here. Such an important analytical parameter as the upper detection limit was dramatically changed by the formation of a polypyrrole conducting polymer layer by the GOx-induced polymerization of polypyrrole (Ppy). An increase in the upper detection limits for differently modified electrodes was estimated by calculation of the apparent Michaelis–Menten constant [KM(app)]. A significant increase in the long-term stability of the GOx-based electrode modified by Ppy (GOx/Ppy) was detected compared with that of an unmodified one. Further application of this approach, based on the self-encapsulation of glucose oxidase and other oxidases, is predicted for such biosensors where extension of the detection rate as well as KM(app)are required. [ABSTRACT FROM AUTHOR]