Estibalitz Ochoteco

Nanostructured disposable impedimetric sensors as tools for specific biomolecular interactions: sensitive recognition of concanavalin A.

The development of sensors to detect specific weak biological interactions is still today a challenging topic. Characteristics of carbohydrate-protein (lectin) interactions include high specificity and low affinity. This work describes the development of nanostructured impedimetric sensors for the detection of concanavalin A (Con A) binding to immobilized thiolated carbohydrate derivatives (D-mannose or D-glucose) onto screen-printed carbon electrodes (SPCEs) modified with gold nanoparticles. Thiolated D-galactose derivative was employed as negative control to evaluate the selectivity of the proposed methodology. After binding the thiolated carbohydrate to the nanostructured SPCEs, different functionalized thiols were employed to form mixed self-assembled monolayers (SAM). Electrochemical impedance spectroscopy (EIS) was employed as a technique to evaluate the binding of Con A to selected carbohydrates through the increase of electron transfer resistance of the ferri/ferrocyanide redox probe at the differently SAM modified electrodes. Different variables of the assay protocol were studied in order to optimize the sensor performance. Selective Con A determinations were only achieved by the formation of mixed SAMs with adequate functionalized thiols. Important differences were obtained depending on the chain lengths and functional groups of these thiols. For the 3-mercapto-1-propanesulfonate mixed SAMs, the electron transfer resistance varied linearly with the Con A concentration in the 2.2-40.0 μg mL(-1) range for D-mannose and D-glucose modified sensors. Low detection limits (0.099 and 0.078 pmol) and good reproducibility (6.9 and 6.1%, n=10) were obtained for the D-glucose and D-mannose modified sensors, respectively, without any amplification strategy.

Screen-printed poly(3,4-ethylenedioxythiophene) (PEDOT): A new electrochemical mediator for acetylcholinesterase-based biosensors

This work describes the use of a PEDOT:PSS-based conductive polymer for designing AChE-based biosensors. The transducers were obtained directly by screen-printing a PEDOT:PSS suspension on the surface of thick film carbon electrodes. The obtained working electrodes showed a high conductivity when compared with electrodes modified with conventional mediators like cobalt phthalocyanine or tetracyanoquinodimethane. The PEDOT:PSS polymer was shown to be suitable for thiocholine oxidation, allowing the measurement of AChE activity at 100 mV vs Ag/AgCl. The high conductivity of PEDOT:PSS allowed the accurate detection of the organophosphate insecticide chlorpyrifos-oxon at concentrations as low as 4x10(-9)M, corresponding to an inhibition ratio of 5%.

Disposable amperometric biosensor based on lactate oxidase immobilised on platinum nanoparticle-decorated carbon nanofiber and poly(diallyldimethylammonium chloride) films.

A novel biosensor for lactate has been developed, using screen-printed carbon electrodes (SPCE) and lactate oxidase (LOx). The active surface of the electrodes was modified using a dispersion of platinum nanoparticle decorated carbon nanofibers (PtNp-CNF) in poly(diallyldimethylammonium) chloride (PDDA) solution. In this way, sensitive, disposable, low cost and reliable hydrogen peroxide sensors were obtained. The immobilisation of LOx on top of these PtNp-CNF-PDDA/SPCEs resulted in amperometric biosensors with high operational stability. The sensitivity of the optimised lactate biosensor was 36.8 (mA/Mcm(2)) with a linear range of 25-1500 µM. The limit of detection was 11 µM (S/N=3). Reproducibility, selectivity and storage stability were also evaluated. Additionally, the stability of the biosensor was also predicted by a model based on thermal degradation. Finally, lactate in sweat and blood samples was determined in a sport test using LOx/PtNp-CNF-PDDA/SPCEs and commercial biosensors respectively. Based on these data, the validity of the sweat lactate for the determination of the lactate threshold is discussed.

Nanotechnology: A Tool for Improved Performance on Electrochemical Screen-Printed (Bio)Sensors

Screen-printing technology is a low-cost process, widely used in electronics production, especially in the fabrication of disposable electrodes for (bio)sensor applications. The pastes used for deposition of the successive layers are based on a polymeric binder with metallic dispersions or graphite, and can also contain functional materials such as cofactors, stabilizers and mediators. More recently metal nanoparticles, nanowires and carbon nanotubes have also been included either in these pastes or as a later stage on the working electrode. This review will summarize the use of nanomaterials to improve the electrochemical sensing capability of screen-printed sensors. It will cover mainly disposable sensors and biosensors for biomedical interest and toxicity monitoring, compiling recent examples where several types of metallic and carbon-based nanostructures are responsible for enhancing the performance of these devices.

Gold coated ferric oxide nanoparticles based disposable magnetic genosensors for the detection of DNA hybridization processes

In this article, a disposable magnetic DNA sensor using an enzymatic amplification strategy for the detection of specific hybridization processes, based on the coupling of streptavidin-peroxidase to biotinylated target sequences, has been developed. A thiolated 19-mer capture probe was attached to gold coated ferric oxide nanoparticles and hybridization with the biotinylated target was allowed to proceed. Then, a streptavidin-peroxide was attached to the biotinylated target and the resulting modified gold coated ferric oxide nanoparticles were captured by a magnetic field on the surface of a home-made carbon screen printed electrode (SPE). Using hydroquinone as a mediator, a square wave voltammetric procedure was chosen to detect the hybridization process after the addition of hydrogen peroxide. Different aspects concerning the assay protocol and nanoparticles fabrication were optimized in order to improve the sensitivity of the developed methodology. A low detection limit (31 pM) with good stability (RSD=7.04%, n=10) was obtained without the need of polymerase chain reaction (PCR) amplification.

Oxireductases in the Enzymatic Synthesis of Water-Soluble Conducting Polymers

This chapter reviews recent advances in the field of biocatalytic synthesis of water-soluble conducting polymers. Biocatalysis is proposed as a versatile tool for synthesis of conducting polymers. First, the enzymatic synthesis of conducting polymers and its mechanism is discussed as well as the use of different type of enzymes. Next, we describe the use of a new bifunctional template (sodium dodecyl diphenyloxide disulfonate) in the synthesis of polyaniline as a strategy to improve the water solubility and electrical conductivity in the obtained polymer. The recent development of enzyme-catalyzed polymerization of 3,4-ethylenedioxythiophene (EDOT) in the presence of polystyrenesulfonate is discussed. This method results in PEDOT materials that show an electrical conductivity of \(2 \times 1{0}^{-3}\,{\mathrm{S\ cm}}^{-1}\) and posses excellent film formation ability, as confirmed by atomic force microscopy images. Finally, a simple method for immobilizing horseradish peroxidases in the biocatalytic synthesis of water-soluble conducting polymers is presented. This method is based on a biphasic catalytic system in which the enzyme is encapsulated inside the ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate, while other components remain in the aqueous phase. The enzyme is easily recovered after reaction and can be reused several times.

Highly Sensitive Detection of Organophosphate Insecticides Using Biosensors Based on Genetically Engineered Acetylcholinesterase and Poly(3,4-Ethylenedioxythiophene)

A poly(3,4-ethylenedioxythiophene) (PEDOT) conducting ink is presented as a new electroactive material to be incorporated in acetylcholinesterase-(AChE-) based screen printed biosensors, acting not only as a conducting template but also as an electrochemical mediator for thiocholine oxidation. Two different strategies have been studied for the chemical synthesis of PEDOT: (a) a classical oxidative polymerisation and (b) a more innovative enzymatic polymerisation, giving a water-soluble PEDOT. The use of this water-soluble conducting polymer as mediator in screen-printed biosensors enables its deposition by printing like the rest of the layers. Highly sensitive acetylcholinesterase-(AChE-) based screen-printed biosensors have been constructed using both classical and enzymatic PEDOT, in combination with genetically modified AChE. These electrodes allow the measurement of thiocholine oxidation at potentials of 100 mV versus Ag/AgCl reference electrode through the mediation of PEDOT. Inhibition of thiocholine production in presence of CPO allow for detection of this pesticide in concentrations as low as 1·10−10 M.

Experimental evaluation of the incidence of tactile sensor limitations on application parameters

This paper presents results from many experiments whose purpose is evaluating the effect of the limitations of a tactile sensor on the tactile image as it is seen for control. Specifically, the variations of a few significant parameters of the tactile image are measured when hysteresis, drift and spatial resolution are taken into account.

Tactile sensors based on conductive polymers

This paper presents results from a few tactile sensors we have designed and fabricated. These sensors are based on a common approach that consists of placing a sheet of piezoresistive material on the top of a set of electrodes. If a force is exerted against the surface of the so obtained sensor, the contact area between the electrodes and the piezoresistive material changes. Therefore, the resistance at the interface changes. This is exploited as transconduction principle to measure forces and build advanced tactile sensors. For this purpose, we use a thin film of conductive polymers as the piezoresistive material. Specifically, a conductive water-based ink of these polymers is deposited by spin coating on a flexible plastic sheet, giving as a result a smooth, homogeneous and conducting thin film on it. The main interest in this procedure is it is cheap and it allows the fabrication of flexible and low cost tactile sensors. In this work we present results from sensors made with two technologies. First, we have used a Printed Circuit Board technology to fabricate the set of electrodes and addressing tracks. Then we have placed the flexible plastic sheet with the conductive polymer film on them to obtain the sensor. The result is a simple, flexible tactile sensor. In addition to these sensors on PCB, we have proposed, designed and fabricated sensors with a screen printing technology. In this case, the set of electrodes and addressing tracks are made by printing an ink based on silver nanoparticles. There is a very interesting difference with the other sensors, that consists of the use of an elastomer as insulation material between conductive layers. Besides of its role as insulator, this elastomer allows the modification of the force versus resistance relationship. It also improves the dynamic response of the sensor because it implements a restoration force that helps the sensor to relax quicker when the force is taken off.

Finite element analysis of tactile sensors made with screen printing technology

Tactile sensors have increasing presence in different applications, especially in assistive robotics or medicine and rehabilitation. They are basically an array of force sensors (tactels) and they are intended to emulate the human skin. Large sensors must be implemented with large area oriented technologies like screen printing. The authors have proposed and made some piezoresistive sensors with this technology. They consist of a few layers of conductive tracks to implement the electrodes and elastomers to insulate them, on a polymer substrate. Another conductive sheet is placed atop the obtained structure. Pressure distribution in the interface between this conductive sheet and the electrodes has a direct impact on the sensor performance. The mechanical behavior of the layered topology with conductive tracks, elastomers and polymers must be studied. For instance, the authors have observed experimentally the existence of pressure thresholds in the response of their sensors. Finite element simulations with COMSOL explain the reason for such thresholds as well as the dependence of the pressure distribution profile on the properties of the materials and the geometry of the tactel. This paper presents results from these simulations and the main conclusions that can be obtained from them related to the design of the sensor.