Browsing by Author "Collyer, Stuart D."
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Item Open Access Construction and interrogation of enzyme microarrays using scanning electrochemical microscopy - optimisation of adsorption and determination of enzymatic activity.(Royal Society of Chemistry, 2011-12-21T00:00:00Z) Roberts, William St John; Davis, Frank; Collyer, Stuart D.; Higson, Seamus P. J.Scanning electrochemical microscopy (SECM) has been used to image and study the catalytic activity of horseradish peroxidase (HRP) immobilised in a patterned fashion onto glass slides. Microarrays of HRP islands could be deposited on amino-modified glass slides using glutaraldehyde crosslinking combined with the SECM being used as a micro-deposition device. The enzymatic activity of the immobilised enzyme on the surface was in the presence of its substrate observed to give rise to substantial positive feedback between the slide and the SECM microelectrode tip. Conversely when either blank slides - or slides coated with HRP which had been subsequently thermally denatured were utilised, these showed negative feedback effects. Various conditions such as enzyme concentration, incubation time and substrate concentration were systematically varied to optimise sensitivity. Regular arrays of HRP could be assembled and when imaged, displayed lower limits of detection of 1.2 × 10(-12) mol ml(-1) of benzoquinone.Item Open Access Electrochemical detection of TNT at cobalt phthalocyanine mediated screen-printed electrodes and application to detection of airborne vapours(Wiley-Blackwell, 2013-10-11) Caygill, J. S.; Collyer, Stuart D.; Holmes, Joanne L.; Davis, Frank; Higson, Seamus P. J.We describe the use of cobalt phthalocyanine as a mediator to improve the sensitivity for the electrochemical detection of TNT. Commercial screen-printed electrodes containing cobalt phthalocyanine were employed for determination of TNT. Improved sensitivities compared to screen-printed carbon electrodes without phthalocyanine were observed, current response for cyclic voltammetric measurements at modified electrodes being at least double that of unmodified electrodes. A synergistic effect between oxygen and TNT reduction was also observed. Correlation between TNT concentrations and sensor output was observed between 0–200 µM TNT. Initial proof-of-concept experiments combining electrochemical determinations, with the use of an air-sampling cyclone, are also reported.Item Open Access The electrochemistry of the ferri/ferrocyanide couple at a calix [4]resorcinarenetetrathiol-modified gold electrode as a study of novel electrode modifying coatings for use within electro-analytical sensors(Elsevier Science B.V., Amsterdam., 2003-06-05T00:00:00Z) Collyer, Stuart D.; Davis, Frank; Lucke, Andrew; Stirling, Charles J. M.; Higson, Seamus P. J.The electrochemistry of the ferri/ferrocyanide redox couple has been studied at Au electrodes modified with calix[4]resorcinarenetetrathiol. Cyclic voltammetry in Fe(CN)63- solutions yields three separate pairs of faradaic peaks. Evidence is given for these redox couples corresponding to the reduction of Fe(CN)63- and the subsequent re-oxidation of Fe(CN)64- in three differing steric arrangements. One pair of peaks suggest that when the Fe(CN)63- ion resides within the calix [4]resorcinarene bowl, electron transport is facilitated by the calix [4]resorcinarene acting as a charge transfer mediator; in this arrangement the activation energy is found to be lowered by ~24kJmol-1. Another pair of peaks is thought to correspond to the reduction of Fe(CN)63- as it approaches the Au electrode by packing itself in-between adjacent calix[4]resorcinarene molecules. The third pair of redox peaks is attributed to the reduction and subsequent re- oxidation of Fe(CN)63-/Fe(CN)64- when the ion resides above a saturated calix [4]resorcinarene coating; in this case the activation energy was raised by ~45kJmol-1. FTIR spectroscopy of calix[4]resorcinarene-coated Au electrodes and calix[4]resorcinarene-coated Au electrodes exposed to Fe(CN)63- lends further support to this argument, by demonstrating that the Fe(CN)63- ion resides within at least two and possibly three differing environments. Calix[4]resorcinarene modified electrodes previously exposed to ferricyanide lose the calix [4]resorcinarene coating together with a surface layer of gold when subsequently scanned in a phosphate buffer. It therefore appears that the calix [4]resorcinarene/Fe(CN)63- association is stronger than the Au binding to the underlying glass material.Item Open Access Flexible Ultrathin PolyDVB/EVB Composite Membranes for the Optimization of a Lactate Sensor.(John Wiley & Sons, Ltd, 2007-01-01T00:00:00Z) Bridge, Kerry; Davis, Frank; Collyer, Stuart D.; Higson, Seamus P. J.An ultrathin composite membrane has been developed as the outer covering barrier in a model amperometric lactate oxidase enzyme electrode. The membrane was formed by cathodic electropolymerization of divinylbenzene/ethylvinylbenzene at the surface of a gold coated polyester support membrane. Permeability coefficients were determined for O2 and lactate across membranes with a range of polymer thicknesses. Anionic interferents (such as ascorbate) were screened from the working electrode by the composite membrane. The composite enzyme electrode showed an increased working concentration range and extended linearity of responses in comparison to an uncoated enzyme electrode.Item Open Access Labeless AC impedimetric antibody-based sensors with pg ml-1 sensitivities for point-of-care biomedical applications.(Elsevier Science B.V., Amsterdam., 2009-01-01T00:00:00Z) Barton, Andrew C.; Collyer, Stuart D.; Davis, Frank; Garifallou, Goulielmos-Zois; Tsekenis, Georgios; Tully, Elizabeth; O'Kennedy, Richard; Gibson, Tim; Millner, Paul A.; Higson, Seamus P. J.This paper describes the development and characterisation of labeless immunosensors for (a) the cardiac drug digoxin and (b) bovine serum albumin (BSA). Commercial screen-printed carbon electrodes were used as the basis for the sensors. Two methods were used to immobilise antibodies at the electrode surface. Aniline was electropolymerised onto these electrodes to form a thin planar film of conductive polyaniline; the polyaniline film was then utilised as a substrate to immobilise biotinylated anti-digoxin using a classical avidin- biotin affinity approach. As an alternative approach, poly(1,2-diaminobenzene) was electrodeposited onto the carbon electrodes and this modified surface was then sonochemically ablated to form an array of micropores. A second electropolymerisation step was then used to co-deposit conductive polyaniline along with antibodies for BSA within these pores to produce a microarray of polyaniline protrusions with diameters of several μm, containing entrapped anti- BSAItem Open Access A new application of scanning electrochemical microscopy for the label-free interrogation of antibody-antigen interactions.(Elsevier Science B.V., Amsterdam., 2011-03-18T00:00:00Z) Holmes, Joanne L.; Davis, Frank; Collyer, Stuart D.; Higson, Seamus P. J.Within this work we present a 'proof of principle' study for the use of scanning electrochemical microscopy (SECM) to detect and image biomolecular interactions in a label-free assay as a potential alternative to current fluorescence techniques. Screen-printed carbon electrodes were used as the substrate for the deposition of a dotted array, where the dots consist of biotinylated polyethyleneimine. These were then further derivatised, first with neutravidin and then with a biotinylated antibody to the protein neuron specific enolase (NSE). SECM using a ferrocene carboxylic acid mediator showed clear differences between the array and the surrounding unmodified carbon. Imaging of the arrays before and following exposure to various concentrations of the antigen showed clear evidence for specific binding of the NSE antigen to the antibody derivatised dots. Non-specific binding was quantified. Control experiments with other proteins showed only non-specific binding across the whole of the substrate, thereby confirming that specific binding does occur between the antibody and antigen at the surface of the dots. Binding of the antigen was accompanied by a measured increase in current response, which may be explained in terms of protein electrostatic interaction and hydrophobic interactions to the mediator, thereby increasing the localised mediator flux. A calibration curve was obtained between 500 fg mL(-1) to 200 pg mL(-1) NSE which demonstrated a logarithmic relationship between the current change upon binding and antigen concentration without the need for any labelling of the substrate.Item Open Access Sonochemically fabricated microelectrode arrays for biosensors offering widespread applicability. Part I(Elsevier Science B.V., Amsterdam., 2004-09-15T00:00:00Z) Barton, Andrew C.; Collyer, Stuart D.; Davis, Frank; Gornall, Davinia D.; Law, Karen A.; Lawrence, Emma C. D.; Mills, Daniel W.; Myler, Suzy; Pritchard, Jeanette A.; Thompson, Mark; Higson, Seamus P. J.A novel and patented procedure is described for the sonochemical fabrication of a new class of microelectrode array based sensor with electrode element populations of up to 2 x 105 cm-2. For some years it has been accepted that microelectrode arrays offer an attractive route for lowering minimum limits of detection and imparting stir (convectional mass transport) independence to sensor responses; despite this no commercial biosensors, to date, have employed microelectrode arrays, largely due to the cost of conventional fabrication routes that have not proved commercially viable for disposable devices. Biosensors formed by our sonochemical approach offer unrivalled sensitivity and impart stir independence to sensor responses. This format lends itself for mass fabrication due to the simplicity and inexpensiveness of the approach; in the first instance impedimetric and amperometric sensors are reported for glucose as model systems. Sensors already developed for ethanol, oxalate and a number of pesticide determinations will be reported in subsequent publications.Item Open Access Sonochemically fabricated microelectrode arrays for biosensors. Part II. Modification with a polysiloxane coating(Elsevier Science B.V., Amsterdam., 2004-09-15T00:00:00Z) Myler, Suzy; Davis, Frank; Collyer, Stuart D.; Higson, Seamus P. J.A polymer modified sonochemically fabricated glucose oxidase microelectrode array with microelectrode population densities of up to 2.5 x 105 microelectrodes cm-2 is reported. These microelectrode sensors were formed by first depositing an insulating film on commercial screen printed electrodes which was subsequently sonicated to form cavities of regular sizes in the film. Electropolymerisation of aniline at the microelectrode cavities formed polyaniline protrusions containing entrapped glucose oxidase. Chemical deposition of polysiloxane from dichlorodimethysilane was used to deposit a thin protective and diffusion mass transport controlling coating over the electrodes. The physical and electrochemical properties of these films were studied. The performance of the final glucose oxidase based microelectrode sensor array is reported.