Urszula Salaj-Kosla

Self-powered wireless carbohydrate/oxygen sensitive biodevice based on radio signal transmission

Here for the first time, we detail self-contained (wireless and self-powered) biodevices with wireless signal transmission. Specifically, we demonstrate the operation of self-sustained carbohydrate and oxygen sensitive biodevices, consisting of a wireless electronic unit, radio transmitter and separate sensing bioelectrodes, supplied with electrical energy from a combined multi-enzyme fuel cell generating sufficient current at required voltage to power the electronics. A carbohydrate/oxygen enzymatic fuel cell was assembled by comparing the performance of a range of different bioelectrodes followed by selection of the most suitable, stable combination. Carbohydrates (viz. lactose for the demonstration) and oxygen were also chosen as bioanalytes, being important biomarkers, to demonstrate the operation of the self-contained biosensing device, employing enzyme-modified bioelectrodes to enable the actual sensing. A wireless electronic unit, consisting of a micropotentiostat, an energy harvesting module (voltage amplifier together with a capacitor), and a radio microchip, were designed to enable the biofuel cell to be used as a power supply for managing the sensing devices and for wireless data transmission. The electronic system used required current and voltages greater than 44 µA and 0.57 V, respectively to operate; which the biofuel cell was capable of providing, when placed in a carbohydrate and oxygen containing buffer. In addition, a USB based receiver and computer software were employed for proof-of concept tests of the developed biodevices. Operation of bench-top prototypes was demonstrated in buffers containing different concentrations of the analytes, showcasing that the variation in response of both carbohydrate and oxygen biosensors could be monitored wirelessly in real-time as analyte concentrations in buffers were changed, using only an enzymatic fuel cell as a power supply.

Characterization of Nanoporous Gold Electrodes for Bioelectrochemical Applications

The high surface areas of nanostructured electrodes can provide for significantly enhanced surface loadings of electroactive materials. The fabrication and characterization of nanoporous gold (np-Au) substrates as electrodes for bioelectrochemical applications is described. Robust np-Au electrodes were prepared by sputtering a gold-silver alloy onto a glass support and subsequent dealloying of the silver component. Alloy layers were prepared with either a uniform or nonuniform distribution of silver and, post dealloying, showed clear differences in morphology on characterization with scanning electron microscopy. Redox reactions under kinetic control, in particular measurement of the charge required to strip a gold oxide layer, provided the most accurate measurements of the total electrochemically addressable electrode surface area, A(real). Values of A(real) up to 28 times that of the geometric electrode surface area, A(geo), were obtained. For diffusion-controlled reactions, overlapping diffusion zones between adjacent nanopores established limiting semi-infinite linear diffusion fields where the maximum current density was dependent on A(geo). The importance of measuring the surface area available for the immobilization was determined using the redox protein, cyt c. The area accessible to modification by a biological macromolecule, A(macro), such as cyt c was reduced by up to 40% compared to A(real), demonstrating that the confines of some nanopores were inaccessible to large macromolecules due to steric hindrances. Preliminary studies on the preparation of np-Au electrodes modified with osmium redox polymer hydrogels and Myrothecium verrucaria bilirubin oxidase (MvBOD) as a biocathode were performed; current densities of 500 μA cm(-2) were obtained in unstirred solutions.

Direct electron transfer of trametes hirsuta laccase adsorbed at unmodified nanoporous gold electrodes

The enzyme Trametes hirsuta laccase undergoes direct electron transfer at unmodified nanoporous gold electrodes, displaying a current density of 28μA/cm(2). The response indicates that ThLc was immobilised at the surface of the nanopores in a manner which promoted direct electron transfer, in contrast to the absence of a response at unmodified polycrystalline gold electrodes. The bioelectrocatalytic activity of ThLc modified nanoporous gold electrodes was strongly dependent on the presence of halide ions. Fluoride completely inhibited the enzymatic response, whereas in the presence of 150mM Cl(-), the current was reduced to 50% of the response in the absence of Cl(-). The current increased by 40% when the temperature was increased from 20°C to 37°C. The response is limited by enzymatic and/or enzyme electrode kinetics and is 30% of that observed for ThLc co-immobilised with an osmium redox polymer.

The spatial and sequential immobilisation of cytochrome c at adjacent electrodes

Two adjacent electrode surfaces were modified in a sequential manner with self-assembled thiol layers from the same solution using conditions (aqueous buffer at neutral pH) suitable for applications with proteins. A faradaic response was obtained from the redox protein, cytochrome c, independently immobilised at each surface.

A pH sensing system using fluorescence-based fibre optical sensor capable of small volume sample measurement

In this work, a low cost optical pH sensing system that allows for small volume sample measurements was developed. The system operates without the requirement of laboratory instruments (e.g. laser source, spectrometer and CCD camera), this lowers the cost and enhances the portability. In the system, an optical arrangement employing a dichroic filter was used which allows the excitation and emission light to be transmitted using a single fibre thus improving the collection efficiency of the fluorescence signal and also the ability of inserting measurement. The pH sensor in the system uses bromocresol purple as the indicator which is immobilised by sol-gel technology through a dip-coating process. The sensor material was coated on the tip of a 1 mm diameter optical fibre which makes it possible for inserting into very small volume samples to measure the pH. In the system, a LED with a peak emission wavelength of 465 nm is used as the light source and a silicon photo-detector is used to detect the uorescence signal. Optical filters are applied after the LED and in front of the photo-detector to separate the excitation and emission light. The fluorescence signal collected is transferred to a PC through a DAQ and processed by a Labview-based graphic-user-interface (GUI). Experimental results show that the system is capable of sensing pH values from 5.3 to 8.7 with a linear response of R2=0.969. Results also show that the response times for a pH changes from 5.3 to 8.7 is approximately 150 s and for a 0.5 pH changes is approximately 50 s.