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A sandwich-type nanostructured immunosensor based on carboxylated multi-walled carbon nanotube (CMWCNT)-embedded whiskered nanofibres (WNFs) was developed for detection of cardiac Troponin I (cTnI). WNFs were directly fabricated on glassy carbon electrodes (GCE) by removing the sacrificial component (polyethylene glycol, PEG) after electrospinning of polystyrene/CMWCNT/PEG nanocomposite nanofibres, and utilised as a transducer layer for enzyme-labeled amperometric immunoassay of cTnI. The whiskered segments of CMWCNTs were activated and utilised to immobilise anti-cTnT antibodies. It was observed that the anchored CMWCNTs within the nanofibres were suitably stabilised with excellent electrochemical repeatability. A sandwich-type immuno-complex was formed between cTnI and horseradish peroxidase-conjugated anti-cTnI (HRP-anti-cTnI). The amperometric responses of the immunosensor were studied using cyclic voltammetry (CV) through an enzymatic reaction between hydrogen peroxide and HRP conjugated to the secondary antibody. The nanostructured immunosensor delivered a wide detection range for cTnI from the clinical borderline for a normal person (0.5-2 ng mL(-1)) to the concentration present in myocardial infarction patients (amp;gt; 20 ng mL(-1)), with a detection limit of similar to 0.04 ng mL(-1). It also showed good reproducibility and repeatability for three different cTnI concentration (1, 10 and 25 ng mL(-1)) with satisfactory relative standard deviations (RSD). Hence, the proposed nanostructured immunosensor shows potential for point-of-care testing., Funding Agencies|Ministry of Science, Research and Technology of Iran; European Commission [PIEF-GA-2013.629251]

The present study describes the development of materials suitable for environmental control of algae. Molecularly imprinted polymers (MIPs) were used as simazine carriers able to provide the controlled release of simazine into water. Three polymers were designed using computational modelling. The selection of methaerylic acid (MA) and hydroxyethyl methacrylate (HEM) as functional monomers was based on results obtained using the Leapfrog algorithm. A cross-linked polymer made without functional monomers was also prepared and tested as a control. The release of simazine from all three polymers was studied. It was shown that the presence of functional monomers is important for polymer affinity and for controlled release of herbicide. The speed of release of herbicide correlated with the calculated binding characteristics. The high-affinity MA-based polymer released similar to 2% and the low-affinity HEM-based polymer released similar to 27% of the template over 25 days. The kinetics of simazine release from HEM-based polymer show that total saturation of an aqueous environment could be achieved over a period of 3 weeks and this corresponds to the maximal simazine solubility in water. The possible use of these types of polymers in the field of controlled release is discussed. (c) 2005 Elsevier B.V. All rights reserved.

A new simple method for the post-polymerisation treatment of molecularly imprinted polymers was proposed. A layer of mineral oil was deposited onto the surface of the polymer in order to create a hydrophobic environment in the binding sites and to improve the recognition properties of the polymer in polar solvents. The testing of polymers performed in acetonitrile showed that the modified polymers possessed significantly increased selectivity as compared with non-treated ones. The three-fold improvement of recognition of the template (cocaine) was achieved; the same time, for non-specific molecule (morphine) the improvement was only 1.3 times. The investigation of the stability of mineral oil coating on the polymer surface suggested that the effect produced is stable over a long period of time. This approach could be used to broaden the range of experimental conditions where molecularly imprinted polymers can perform successfully. (c) 2005 Elsevier B.V. All rights reserved.

Recently, the development and optimization of a flow injection analysis (FIA) operated bacterial biosensor based on the aerobic catabolism of Pseudomonas putida ML2 was reported in the literature (Lanyon et al. 2004, 2005). By adapting information from these reports, we investigated whether operating parameters and procedures of the benzene biosensor could be directly applied to a new system based on a different bacterial strain for the detection of the whole benzene, toluene, ethylbenzene, and xylenes range. Cells of the investigated bacterial strain, Pseudomonas putida F1, were immobilized between two cellulose acetate membranes and fixed onto a Clark dissolved oxygen electrode. The P. putida F1 aerobically degrades benzene, toluene, and ethylbenzene (BTE) (Cho et al. 2000). The BTE biosensor in kinetic mode FIA displayed a linear range of 0.02-0.14 mM benzene (response time: 5 min, baseline recovery time: 15 min), 0.05-0.2 mM toluene (response time: 8 min, baseline recovery time: 20 min), and 0.1-0.2 mM ethylbenzene (response time: 12 min, baseline recovery time: 30 min), respectively. Due to the differences in sensitivity, response, and baseline recovery times for BTE, it was possible to differentiate each compound in mixtures of these volatile organic compounds (VOCs). No response for xylenes could be obtained since they cannot be completely metabolized by this bacterial strain. However, it was reported that the range of compounds degradable by P. putida F1 can possibly be expanded by cultivating the cells on different carbon sources (Choi et al. 2003). The sensor showed good intra- and interassay reproducibility, and all obtained results were comparable with those reported in the literature. The demonstrated reproducibility and the simplicity and ease of use as well as the portability for in situ measurements indicates that the biosensor could be suitable as a reliable initial warning device for elevated BTE levels in indoor and outdoor environments.

Twenty years on from a review in the first issue of this journal, this contribution revisits glucose sensing for diabetes with an emphasis on commercial developments in the home blood glucose testing market. Following a brief introduction to the needs of people with diabetes, the review considers defining technologies that have enabled the introduction of commercial products and then reviews the products themselves. Drawing heavily on the performance of actual instruments and publicly available information from the companies themselves, this work is designed to complement more conventional reviews based on papers published in scholarly journals. It focuses on the commercial reality today and the products that we are likely to see in the near future. (c) 2004 Elsevier B. V. All rights reserved.

A DNA-based surface plasmon resonance (SPR) biosensor has been developed for the detection of TP53 mutation using the inexpensive and commercially available instrument, SPREETA (TM) SPR-EVM-BT, from Texas Instruments. A direct immobilisation procedure, based on the coupling of thiol-derivatised oligonucleotide probes (Probe-C6-SH) to bare gold sensor surfaces, was optimized using synthetic oligonucleotides. Hybridisation reactions between the immobilised probe and a short sequence (26 mer) complementary, non-complementary and one-point mutation DNA were then investigated. The main analytical parameters of the sensor system were studied in detail including selectivity, sensitivity, reproducibility and analysis time. Finally, the sensor system was successfully applied to polymerase chain reaction (PCR)-amplified real samples, DNA extracted from both normal, wild-type, (Jurkat) and mutated (Molt 4), carrying the mutation at codon 248 of the TP53 cell lines. The results obtained demonstrate that the DNA-based SPR biosensor was able to distinguish sequences present in the various samples that differ only by one base; and hence, it appears to be a strong candidate technique for the detection of gene mutation. (c) 2004 Elsevier B.V. All rights reserved.

A set of polymers was imprinted with (-)-ephedrine using thermal and UV initiation. The performance of the synthesized materials was studied by HPLC. It was shown that the polymer morphology and enantioselective properties are dependent on the polymerization conditions and time of the reaction. The binding mechanism of synthesized polymers was studied using Vant Hoff analysis. The results of testing strongly indicate that the polymer receptor structure is shaped during the initial phase of polymer gelation. The present study shows the importance of controlling experimental conditions in the MIP synthesis and highlights potential problems expected during scaling up of MIP production.

Tissue engineering is one of the most exciting and rapidly growing areas in biomedical engineering that offers vast potential for changing traditional approaches to meeting many pharmaceutics and critical health care needs. Currently the bottle-neck area in this multidisciplmiary field appears to be materials and fabrication technology for the design of artificial extracellular matrices/scaffolds that support culturing and growth of new tissue. We have shown that stable relief structures can be created and maintained in the bulk of ice by continuous s canning with computer-guided IR CO2 laser. The optimal laser beam intensity and fluence rate distribution within the ice sample, as well as the rate of scanning were estimated based on the Monte Carlo model utilized physical/optical properties of ice. The results of numerical simulation are agreed well with the observed experimental results of thermo-coupling measurements and obtained microscopic images.

Allergen levels in indoor environments, leading to many diseases, eg asthma, rhinitis and conjunctivitis, affect a large and increasing fraction of the population. A quite effective and inexpensive method of a rough but very rapid overall assessment of total allergen level in the environment has been developed. The method involved estimation of protein in allergen extracts by screen-printed electrodes using two different techniques. The biosensor comprised a rhodinised carbon working electrode, a silver/silver chloride reference electrode and a carbon counter electrode. In the first method the enzyme protease reacted with allergen protein to release amino acid, which produced hydrogen peroxide in the presence of amino acid oxidase. This was detected amperometrically. The second method used potassium bromide as electrolyte and the electrode was subjected to dual potential. Bromine, released due to electrolysis at higher potential, was consumed by the allergen protein at lower potential. In the first method, a unique technique was used to microencapsulate the enzyme protease and immobilise it on the surface of the electrode by in-situ polymerisation to avoid contact with the amino acid oxidase. A total of seven allergens were tested and the results gave a good correlation with the standard protein measurement method. Environmental specimens from indoors, schools and workplaces can be evaluated for the aeroallergens produced by dust mites, animal hairs, cockroach debris, pollens, etc as a means of determining the exposure risk. (c) 2005 Society of Chemical Industry.

A molecularly imprinted polymer (MIP) film for domoic acid (DA) was synthesised by direct photo-grafting onto a gold chip suitable for a surface plasmon resonance (SPR) based bioanalytical instrument system, the BlAcore 3000(TM). The gold surface was first functionalised with a self-assembled monolayer of 2-mercaptoethylamine and subsequent carbodiimide chemistry was performed for covalent attachment of the photoinitiator, 4,4-azobis(cyanovaleric acid). This ensured that the formation of the MIP thin film, comprising 2-(diethyl amino) ethyl methacrylate as functional monomer and ethylene glycol dimethacrylate as cross-linker, occurred only at the surface level. Optimisation and control over the grafting procedure were achieved using contact angle measurements and atomic force microscope (AFM) imaging. The surface grafting resulted in the formation of thin and homogeneous MIP film with thickness of 40 nm. A competitive binding assay was performed with free DA and its conjugate with horseradish peroxidase, which was used as a refractive label. The sensor was evaluated for its sensitivity, cross-reactivity, and robustness by using a BlAcore 3000(TM). Likewise, monoclonal antibodies acting as natural receptors for the toxin were studied with the same BlAcore system. Results of a comparison between the artificial and natural receptors are reported. In contrast to monoclonal antibodies, the regeneration of MIP chip did not affect its recognition properties and continuous measurement was possible over a period of at least 2 months. (C) 2004 Elsevier B.V. All rights reserved.

A novel method of photochemical polymerization of thiophene derivatives in aqueous solution catalyzed by potassium dichromate and initiated by illumination is described.

Genosensor technology relying on the use of carbon and gold electrodes is reviewed. The key steps of each analytical procedure, namely DNA-probe immobilisation, hybridisation, labelling and electrochemical investigation of the surface, are discussed in detail with separate sections devoted to label-free and newly emerging magnetic assays. Special emphasis has been given to protocols that have been used with real DNA samples. (C) 2003 Elsevier B.V. All fights reserved.

A set of polymers was imprinted with (+)-ephedrine at ambient and 10 bar pressure. The performance of the synthesized materials was studied by HPLC at six different temperatures: 0, 10, 20, 30, 40, and 50 degreesC. It was shown that the polymer morphology (surface area and porosity) and enantioselective properties are different for these two polymers-an unexpected result which was in contradiction with the theoretical forecasts. A possible mechanism which explains the observed phenomenon is discussed. The contributing factor which influenced the polymer performance was the polymer swelling. This shows that, in addition to temperature, other physical parameters such as pressure and polymer swelling should be analyzed and accounted for in the quality control of MIP synthesis and application.

Microsystin-LR is one of the most widespread and dangerous toxins produced by the freshwater Cyanobacteria. The contamination of water supplies with microcystin-LR has been reported in several areas around the world and the development of an easy-to-use, rapid, robust and inexpensive sensor for this toxin is urgently required. In this work an artificial receptor for microcystin-LR was synthesised using the technique of molecular imprinting. The composition of the molecularly imprinted polymer (MIP) was optimised using computer modelling. The synthesised polymer was used both as a material for solid-phase extraction (SPE) and as a sensing element in a piezoelectric sensor. Using the combination of SPE followed by detection with a piezoelectric sensor the minimum detectable amount of toxin was 0.35 nM. The use of MIP-SPE provided up to 1000 fold preconcentration, which was more than sufficient for achieving the required detection limit for microcystin-LR in drinking water (I nM). This work is the first example where the same MIP receptor has been used successfully for both SPE and the corresponding sensor. (C) 2002 Elsevier Science B.V. All rights reserved.

Extending the previous preliminary study on the construction of a capillary electrophoresis (CE)/sensor for the detection of reducing analytes, we focus the interest on the simultaneous detection of redox active species, which are important indicators of the oxidative damage in tissues, of food preservation, and of pollution. The CE/sensor was built by modifying the detector-portion of the capillary with the redox-sensitive polymer polyaniline (PANI). The analyte is detected by monitoring the changes in optical absorption of the PANI film. The CE/sensor was tested, with good results, with ascorbic acid, glutathione (GSH), as well as with compounds with very close similarity (ascorbic and isoascorbic acid). The kinetics of oxidation and reduction of PANI were evaluated. Further a PANI/CE-biological sensor was developed by coupling an enzyme, glucose oxidase (GOD), to the PANI-modified portion of the capillary. The stability of the immobilized GOD and the sensitivity of the CE/biosensor were studied, by using glucose as test analyte in concentrations within the physiological range. The results indicate that the CE/biosensor had good stability (more than 75% of original activity retained after 30 operational days), manufacturing reproducibility and a sensing range convenient for monitoring physiological glucose (1-24 mm).

In order to encourage more exposure measurements to be performed, a formic acid gas-phase biosensor has been developed for this purpose. In the present paper, an enzyme based biosensor has been validated with respect to analyte selectivity and on-site use. To ensure that the sampler developed measures the compound of interest the biosensor was exposed to three near structural homologues to formic acid, i.e. acetic acid, methanol and formaldehyde. These vapours were generated with and without formic acid and the only compound that was found to have an effect on the performance of the biosensor, albeit a small one, was acetic acid. The field test was performed in a factory using formic acid-containing glue for glulam products. In parallel to the measurements with the biosensor a well defined reference method was used for sampling and analysing formic acid. It was found that the biosensor worked satisfactorily in this environment when used in a stationary position. It was also shown that the biosensor could determine formic acid vapour concentrations down to 0.03 mg m(-3).

Molecularly imprinted polymer membranes for a model compound, atrazine, were prepared in situ from a monomer mixture containing methacrylic acid, tri(ethylene glycol) dimethacrylate, and atrazine using U-V-initiated polymerization. To improve flexibility and mechanical stability of these membranes, oligourethane acrylate was added to the mixture of monomers. Polymeric additives were used to increase porosity of membranes and their permeability as well as to make them suited for filtration experiments. This process resulted in the formation of thin, flexible, and porous membranes containing atrazine-specific binding sites. The atrazine-imprinted membranes showed higher affinity to this herbicide than to structurally similar compounds (simazine, prometryn, and metribuzin). The binding capacity of MIP membranes was found to be significantly higher than that observed previously for MIP systems. The high affinity, specificity, and binding capacity of MIP membranes, together with their straightforward and easy preparation, provide a good basis for their application in separation and purification, e.g., in membrane chromatography.

Over the past two decades, molecularly imprinted polymers (MIPs) have attracted broad interest from scientists engaged in sensor development. This attention can be explained by the serious potential advantages of using MIPs in pi ace of natural receptors and enzymes such as their superior stability, low cost and easy preparation. This review encompasses recent achievements in molecular imprinting related to the area of sensor technology. Since electrochemical biosensors dominate the market and due to specific requirements of this journal, the emphasis of this review will be on the development of electrochemical MIP sensors. The problems associated with application of imprinted polymers in sensors are highlighted and possible solutions indicated. The commercial potential of MIP-based sensors is analyzed in the expectation that they can offer improved performance in the analytical market place.

Biosensors are analytical devices that use a biological or biologically derived material immobilized at a physicochemical transducer to measure one or more analytes. Although there are a large number of reviews on biosensors in general, there has been little systematic information. presented on the application of natural receptors in sensor technology. This perspective discusses broadly the fundamental properties of natural receptors; which make them an attractive option for use as biorecognition elements in. sensor technology. It analyses the current situation by reference to, typical examples, such as the application of nicotinic acetylcholine receptor and G protein-linked receptors in affinity sensors and analyses the problems that need to be resolved prior to any commercialization of such devices.

An immunosensor for the determination of okadaic acid (OA) using a quartz crystal microbalance (QCM) was developed and optimised in standard solutions. Several coupling techniques, protein A, protein G and polyethylenimine (PEI) with glutaraldehyde (GA) cross-linking, were investigated for the determination of okadaic acid and a very good result was obtained with PEI coupling. With the PEI coupling method, the optimisation of incubation time for the activation of PEI on the crystal surface using GA, the effect of the dilution factor of OA-bovine serum albumin (BSA) conjugate and the amount of antibody on crystal frequency were studied. Different molar ratios (4:1, 14:1, 30:1) of OA to bovine serum albumin for the conjugation were examined and the results using ELISA and a QCM showed that a ratio of 14:1 was slightly better than the other two. The strong attachment of the cross-linked complex to the gold surface resulted in an excellent storage lifetime of 38 days. However, the detection limit (1.9 mug/ml) and the sensitivity of the sensor were not satisfactory. Significant improvement of the performance of the device was obtained by incorporating an antibody-BSA hydrogel. Initial results showed that the minimum amount of analyte detectable and the sensitivity of the device were improved by 524- and 80-fold, respectively. (C) 2002 Elsevier Science B.V. All rights reserved.

Aims: Use of an electronic nose (e.nose) system to differentiation between anaerobic bacteria grown in vitro on agar media. Methods and Results: Cultures of Clostridium spp. (14 strains) and Bacteroides fragilis (12 strains) were grown on blood agar plates and incubated in sampling bags for 30 min before head space analysis of the volatiles. Qualitative analyses of the volatile production patterns was carried out using an e.nose system with 14 conducting polymer sensors. Using data analysis techniques such as principal components analysis (PCA), genetic algorithms and neural networks it was possible to differentiate between agar blanks and individual species which accounted for all the data. A total of eight unknowns were correctly discriminated into the bacterial groups. Conclusions: This is the first report of in vitro complex volatile pattern recognition and differentiation of anaerobic pathogens. Significance and Impact of the Study: These results suggest the potential for application of e.nose technology in early diagnosis of microbial pathogens of medical importance.

Measurement of ecological, climatic and anthropogenic changes underpins the formulation of effective management strategies for sustainable use and protection of the marine environment. Sensors are traditionally used in marine studies to determine physical parameters, but there is increasing demand for real-time information about chemical and biological parameters. These parameters are currently measured in samples collected at sea and subsequently analysed in the laboratory. Biosensors fuse the exquisite sensitivity and specificity of living systems with the processing power of microelectronics to deliver simple, inexpensive measurement systems for use in the field or deployment in situ. While their potential for use in the marine environment is enormous, much published work to date has focussed on applications in freshwater and wastewater. Marine applications pose a substantial challenge in the robustness required for remote application, but recent developments in portable medical devices and receptor design suggest that these demands can now be realistically tackled. (C) 2002 Elsevier Science Ltd. All rights reserved.

A set of polymers has been imprinted with (-)-ephedrine at six different temperatures, ranging from -30 to +80 degreesC. Polymer affinity and specificity were observed to be strongly dependent on the polymerization temperature. The experimental results suggest that the polymer is able to "memorize" the temperature used in the polymerization process in a manner similar to previously documented MIP "memory" effects for the template and polymerization solvent. In a study of the effect of temperature on retention and selectivity in HPLC (using the MIP as a column packing), a clear gradient change in the Vant Hoff plots was observed at 20-30 degreesC. This indicates a transition in binding mechanism from exothermic at higher temperatures to endothermic at lower temperatures. These results, supported by the evidence of template-induced MIP swelling, are interpreted in terms of desolvation and conformational changes in the polymers induced by the interaction with the template.

A computational approach for the design of a molecularly imprinted polymer (MIP) specific for Cyanobacterial toxin microcystin-LR is presented. By using molecular modeling software, a virtual library of functional monomers was designed and screened against the target toxin, employed as a template. The monomers giving the highest binding energy were selected and used in a simulated annealing (molecular dynamics) process to investigate their interaction with the template. The stoichiomettic ratio observed from the simulated annealing study was used in MIP preparation for microcystin-LR. The monomers were copolymerized with a cross-linker in the presence of the template. A control (blank) polymer was prepared under the same conditions but in the absence of template. A competitive assay with microcystin-horseradish peroxidase conjugate was optimized and used to evaluate the affinity and cross-reactivity of the polymer. The performance of the artificial receptor was compared to the performance of monoclonal and polyclonal antibodies raised against the toxin. The results indicate that imprinted polymer has affinity and sensitivity comparable to those of polyclonal antibodies (die detection limit for microcystin-LR using the MIP-based assay was found to be 0.1 mug L-1), while superior chemical and thermal stabilities were obtained. Moreover, cross-reactivity to other toxin analogues was very low for the imprinted polymer, in contrast to the results achieved for antibodies. It is anticipated that the polymer designed could be used in assays, sensors, and solid-phase extraction.

The high sensitivity and specificity of DNA hybridisation techniques makes them powerful tools for environmental or clinical analysis. This work describes the development of a DNA piezoelectric biosensor for the detection of the hybridisation reaction. Attention was focused on the choice of the coating chemistry that could be used for the immobilisation of oligonucleotides onto the gold surface of the quartz crystal. Four immobilisation procedures were tested and compared considering the amount of immobilised probe, the extent of the hybridisation reaction, the possibility of regeneration and the absence of non-specific adsorption. All the experiments were performed with oligonucleotides of 25 bases (probe, target and non-complementary oligonucleotide). The four coating methods were all based on the use of self-assembled monolayers (SAM). Three of them employed the interaction between streptavidin and biotin for the immobilisation of a biotinylated probe. Results indicated that immobilisation of a biotinylated probe on streptavidin linked to a layer of carboxylated dextran provides higher sensitivity for the detection of the hybridisation reaction, absence of non-specific adsorption and a higher stability with respect to the regeneration step. (C) 2002 Elsevier Science B.V. All rights reserved.

The use of volatile production patterns produced by bacterial contaminants in urine samples were examined using electronic nose technology. In two experiments 25 and 45 samples from patients were analysed for specific bacterial contaminants using agar culture techniques and the major UTI bacterial species identified. These samples were also analysed by incubation in a volatile generation test tube system for 4-5 h. The volatile production patterns were then analysed using an electronic nose system with 14 conducting polymer sensors. In the first experiment analysis of the data using a neural network (NN) enabled identification of all but one of the samples correctly when compared to the culture information. Four groups could be distinguished, i.e. normal urine, Escherichia coli infected, Proteus spp. and Staphylococcus spp. In the second experiment it was again possible to use NN systems to examine the volatile production patterns and identify 18 of 19 unknown UTI cases. Only one normal patient sample was mis-identified as an E coli infected sample. Discriminant function analysis also differentiated between normal urine samples, that infected with E coli and with Staphylococcus spp. This study has shown the potential for early detection of microbial contaminants in urine samples using electronic nose technology for the first time. These findings will have implications for the development of rapid systems for use in clinical practice. (C) 2002 Elsevier Science B.V. All rights reserved.

The capillary walls of fused-silica capillary electrophoresis (CE) columns were modified with a thin film of polyaniline (PANI), providing open-tubular columns with a stable coating containing aromatic groups and amine functionalities. Fast and efficient separations were observed for small bioactive peptides under acidic conditions on PANI-coated columns. The mechanism of separation is based on hydrophobic interactions between the analytes and the polymeric matrix. Good reproducibility was observed from run-to-run. Due to the simple derivatization procedure, method flexibility, the uniformity of the coating and its stability, conjugated polymers could find practical application in capillary zone electrophoresis (CZE) separations.

A CE/biosensor for measuring ascorbic acid was developed by coupling a polyaniline optical sensor and capillary electrophoresis (CE). The capillary column was partially modified with a thin film of polyaniline redox sensitive material. Ascorbic acid was detected by monitoring the changes in optical absorbance occurring to the polyaniline film upon the reduction reaction. The sensor response (change in optical absorbance at 650 nm) is proportional to the concentration of ascorbic acid over a range of 2.5-250 mg/L and the response range has shown a clear dependence on the characteristics of the polymerized film. High specificity and sensitivity of the present method, low sample consumption, short times of response (ca. 2 min) and the reproducibility of the results demonstrate that the CE/polyaniline-sensor could be further employed in the study of the relation between the content of L-ascorbic acid in body fluids and clinical parameters, e.g., cell ageing.

Molecular recognition between molecules is one of the most fundamental processes in biology and chemistry. The recognition process is largely driven by non-covalent forces such as hydrogen bonding, electrostatics, van der Waals forces, pi-pi interactions, and conformational energy. The complementarity between the receptor and substrate is very similar to the "lock and key" function, first described by Emil Fischer over 100 years ago, - the lock being the molecular receptor such as a protein or enzyme and the key being the substrate such as a drug, that is recognized to give a defined receptor-substrate complex. This review focuses on the design of specific ligand systems as "Keys" to enable the induced fit of these keys into the target macromolecules, protein/enzyme (Locks) with particular emphasis on protein recognition.

A formic acid biosensor for air monitoring has been evaluated using chemometric methods. Using experimental design eleven factors that could influence the performance of the biosensor were examined. The response matrices consisted of six parameters (steady state currents at three different formic acid concentrations and response rates during changes in formic acid concentrations) describing the performance of the biosensor. The data were evaluated using a combination of principal component analysis (PCA) and multiple linear regression (MLR). To confirm the conclusions from the PCA-MLR partial least squares (PLS) was also used. The most important factor for the biosensor performance was found to be the enzyme concentration. Using the information from the chemometric analyses the optimum operation conditions for the biosensor were determined. The steady state currents were increased by 18-30% and the initial two response rates increased by 47-89% compared with a biosensor that had not been optimised.

A new technique for coating microtitre plates with molecularly imprinted polymers (MIP), specific for low-molecular weight analytes (epinephrine, atrazine) and proteins is presented. Oxidative polymerization was performed in the presence of template; monomers: 3-aminophenylboronic acid (APBA), 3-thiopheneboronic acid (TBA) and aniline were polymerized in water and the polymers were grafted onto the polystyrene surface of the microplates. It was found that this process results in the creation of synthetic materials with antibody-like binding properties. It was shown that the MIP-coated microplates are particularly useful for assay development. The high stability of the polymers and good reproducibility of the measurements make MIP coating an attractive alternative to conventional antibodies or receptors used in enzyme linked immunosorbent assay (ELISA). (C) 2001 Elsevier Science B.V. All rights reserved.

A new type of membrane with potential-controllable functions was prepared by grafting polyaniline to the surface of track-etch Nucleopore membrane with a vacuum deposited gold layer (PANI-membranes). Because of the cylindrical shape of these pores, a nanocylinder of the desired conducting material is obtained in each pore. A potential applied to PANI-membranes governs the properties of the conducting layer, such as charge and its distribution, conformation, and density and, hence, controls the diffusion rate of substances through these membranes. The variation in diffusion rates of different substances with different potentials applied to PANI-membranes provides the basis for the application of these membranes in separation processes and sensors.

A method for the selective detection of creatinine is reported, which is based on the reaction between polymerised hemithioacetal formed by allyl mercaptan, o-phthalic aldehyde, and primary amine leading to the formation Of fluorescent isoindole complex. This method has been demonstrated previously for the detection of creatine using creatine-imprinted molecularly imprinted polymers (MlPs) Since MlPs created using traditional methods were unable to differentiate between creatine and creatinine, a new approach to the rational design of a molecularly imprinted polymer (MIP) selective for creatinine was developed using computer simulation. A virtual library of functional monomers was assigned and screened against the target molecule, creatinine, using molecular modelling software. The monomers giving the highest binding score were further tested using simulated annealing in order to mimic the complexation of the functional monomers with template in the monomer mixture. The result of this simulation gave an optimised MIP composition. The computationally designed polymer demonstrated superior selectivity in comparison to the polymer prepared using traditional approach, a detection limit of 25 muM and good stability. The Bite-and-Switch approach combined with molecular imprinting can be used for the design of assays and sensors, selective for amino containing substances. (C) 2001 Elsevier Science B.V. All rights reserved.

A new approach to the computational design of molecularly imprinted polymers (MIP) specific for ephedrine is presented. A virtual library of functional monomers was developed and screened against the template using molecular modelling software. The monomers giving the highest binding score were co-polymerized with a cross-linker in the presence of ephedrine. Control (blank) polymers were prepared under the same conditions but in the absence of the template. A good correlation was found between the modelling results and performance of the materials in an HPLC study. A MIP based on one of the selected monomers-hydroxyethyl methacrylate-gave a separation of ephedrine enantiomers with a separation factor a of 1.42-2.09 (depending on temperature). This figure is larger than the a values generally obtained with commercially available chiral phases. It is anticipated that the computational approach will be of use for the rational design of MIPS and the prediction of polymer affinity and specificity.

A new polymer capable of reacting with primary amines was synthesised from allyl mercaptan, o-phthalic dialdehyde and ethylene glycol dimethacrylate by radical polymerisation. Reactive hemithioacetal formed by allyl mercaptan and dialdehyde can bind primary amino groups without additional pre-activation forming the fluorescent isoindole complex. It gives a great opportunity to monitor binding and perform loading of the amino compounds onto the reactive surface. The reactive polymer is found to be an effective matrix for immobilisation of the proteins and other amino-containing compounds in affinity chromatography and could be used for their detection in solution. (C) 2001 Elsevier Science Ltd. All rights reserved.

A technique for coating microplate wells with molecularly imprinted polymers (MIPs) specific for proteins is presented. 3-Aminophenylboronic acid was polymerized in the presence of the following templates: microperoxidase, horseradish peroxidase, lactoperoxidase, and hemoglobin, via oxidation of the monomer by ammonium persulfate. This process resulted in the grafting of a thin polymer layer to the polystyrene surface of the microplates. Imprinting resulted in an increased affinity of the polymer toward the corresponding templates. The influence of the washing procedure, template concentration, and buffer pH on the polymer affinity was analyzed. It was shown that the stabilizing function of the support and spatial orientation of the polymer chains and template functional groups are the major factors affecting the imprint formation and template recognition. Easy preparation of the MIPs, their high stability, and their ability to recognize small and large proteins, as well as to discriminate molecules with small variations in charge, make this approach attractive and broadly applicable in biotechnology, assays and sensors.

Molecularly imprinted polymers (MIPs) represent a new class of materials that have artificially created receptor structures(1-3). Since their discovery in 1972, MIPs have attracted considerable interest from scientists and engineers involved with the development of chromatographic adsorbents, membranes, sensors and enzyme and receptor mimics.

A technique for modification of the microtiter reader plates well with a polyaniline (PANI) film sensitive for ascorbic acid is presented. The principle of the analyte detection is based on monitoring the changes in optical absorption of the PANI film resulting from the reduction process initiated by ascorbic acid. The detection limit for ascorbic acid is 1 mg/L. Testing with real samples (soft drinks, fruit juices) gave good correlation of the method with iodimetric titration. High sensitivity, stability, and good reproducibility of the measurements make the proposed system an attractive alternative to traditional assays, used in medicine, ecology and biotechnology.

The detection of creatine is important in the analysis of athletes and body builders. Here is reported the preparation of a synthetic polymer using imprinting polymerization, which leaves the polymer with receptor-mimicking recognition sites that are specific for creatine. Molecular recognition results in a fluorescent complex (see Figure) and thus represents a bite-and-switch" mechanism.

Recently the use of smell in clinical diagnosis has been rediscovered due to major advances in odour sensing technology and artificial intelligence (AI). It was well known in the past that a number of infectious or metabolic diseases could liberate specific odours characteristic of the disease stage. Later chromatographic techniques identified an enormous number of volatiles in human clinical specimens that might serve as potential disease markers. "Artificial nose" technology has been employed in several areas of medical diagnosis, including rapid detection of tuberculosis (TB), Helicobacter pylori (HP) and urinary tract infections (UTI). Preliminary results have demonstrated the possibility of identifying and characterising microbial pathogens in clinical specimens. A hybrid intelligent model of four interdependent "tools", odour generation "kits", rapid volatile delivery and recovery systems, consistent low drift sensor performance and a hybrid intelligent system of parallel neural networks (NN) and expert systems, have been applied in gastric, pulmonary and urine diagnosis. Initial clinical tests have shown that it may be possible in the near future to use electronic nose technology not only for the rapid detection of diseases such as peptic ulceration, UTI, and TB but also for the continuous dynamic monitoring of disease stages. Major advances in information and gas sensor technology could enhance the diagnostic power of future bio-electronic noses and facilitate global surveillance models of disease control and management.