Controlled release of the herbicide simazine from computationally designed molecularly imprinted polymers

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 methacrylic 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 2% and the low-affinity HEM-based polymer released 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 discuss

Selection of imprinted nanoparticles by affinity chromatography

Soluble molecularly imprinted nanoparticles were synthesised via iniferter initiated polymerisation and separated by size via gel permeation chromatography. Subsequent fractionation of these particles by affinity chromatography allowed the separation of high affinity fractions from the mixture of nanoparticles. Fractions selected this way possess affinity similar to that of natural antibodies (Kd 6.6 × 10−8) M and were also able to discriminate between related functional analogues of the templ

Preliminary evaluation of military, commercial and novel skin decontamination products against a chemical warfare agent simulant (methyl salicylate)

This is an Accepted Manuscript of an article published by Taylor & Francis in Cutaneous and Ocular Toxicology on 13 August 2015, available online: http://www.tandfonline.com/doi/full/10.3109/15569527.2015.1072544.Rapid decontamination is vital to alleviate adverse health effects following dermal exposure to hazardous materials. There is an abundance of materials and products which can be utilised to remove hazardous materials from the skin. In this study, a total of 15 products were evaluated, 10 of which were commercial or military products and 5 were novel (molecular imprinted) polymers. The efficacies of these products were evaluated against a 10µL droplet of 14C-methyl salicylate applied to the surface of porcine skin mounted on static diffusion cells. The current UK military decontaminant (Fuller’s earth) performed well, retaining 83% of the dose over 24 hours and served as a benchmark to compare with the other test products. The five most effective test products were Fuller’s earth (the current UK military decontaminant), Fast-Act® and three novel polymers (based on itaconic acid, 2-trifluoromethylacrylic acid and N,N-methylene bis acrylamide). Five products (medical moist free wipes, 5% FloraFree™ solution, normal baby wipes, baby wipes for sensitive skin and Diphotérine™) enhanced the dermal absorption of 14C-methyl salicylate. Further work is required to establish the performance of the most effective products identified in this study against chemical warfare agents.Peer reviewedFinal Accepted Versio

The stabilisation of receptor structure in low cross-linked MIPs by an immobilised template

In molecularly imprinted polymers (MIPs) a high level of cross-linking is usually important for preserving the receptor structure. We propose here an alternative approach for stabilising binding sites, which involves the use of an immobilised template. The idea is based on the assumption that an immobilised template will ‘‘hold’’ polymeric chains and complementary functionalities together, preventing the collapsing of the binding sites. To test this postulate, a range of polymers was prepared using polymerisable (2,4-diamino-6- (methacryloyloxy)ethyl-1,3,5-triazine) and non-polymerisable (or extractable) (2,4-diamino-6-methyl-1,3,5-triazine) templates, methacrylic acid as functional monomer and ethylene glycol dimethacrylate as cross-linker. The level of cross- linking was varied from 12 to 80%. Polymerisations were performed in acetonitrile using UV initiation. Binding properties of the synthesised materials were characterised both by HPLC and equilibrium batch binding experiments followed by HPLC-MS or UV-visible detection. The adsorption isotherms of polymers were obtained and fitted to the Langmuir model to calculate dissociation constant, Kd, and concentration of binding sites for each material. The results strongly indicate that the presence of an immobilised template improves the affinity of MIPs containing low percentages of cross- linker. The low cross-linked MIPs synthesised with a polymerisable template also retain a reasonable degree of selectivity. Low crosslinked MIPs with such binding characteristics would be useful for the creation of new types of optical and electrochemical sensors, where induced fit or the ‘‘gate effect’’ could be used more effectively for generating and enhancin

Substitution of antibodies and receptors with molecularly imprinted polymers in enzyme-linked and fluorescent assays

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, 3- thiopheneboronic acid 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 ELISA

Direct replacement of antibodies with molecularly imprinted polymer (MIP) nanoparticles in ELISA - development of a novel assay for vancomycin

A simple and straightforward technique for coating microplate wells with molecularly imprinted polymer nanoparticles (nanoMIPs) to develop ELISA type assays is presented here for the first time. NanoMIPs were synthesized by a solid phase approach with immobilized vancomycin (template) and characterized using Biacore 3000, dynamic light scattering and electron microscopy. Immobilization, blocking and washing conditions were optimized in microplate format. The detection of vancomycin was achieved in competitive binding experiments with a HRP-vancomycin conjugate. The assay was capable of measuring vancomycin in buffer and in blood plasma within the range 0.001-70 nM with a detection limit of 0.0025 nM (2.5 pM). The sensitivity of the assay was three orders of magnitude better than a previously described ELISA based on antibodies. In these experiments nanoMIPs have shown high affinity and minimal interference from blood plasma components. Immobilized nanoMIPs were stored for 1 month at room temperature without any detrimental effects to their binding properties. The high affinity of nanoMIPs and the lack of a requirement for cold chain logistics make them an attractive alternative to traditional antibodies used in ELIS

Catalytic molecularly imprinted polymer membranes: Development of the biomimetic sensor for phenols detection

Portable biomimetic sensor devices for the express control of phenols content in water were developed. The synthetic binding sites mimicking active site of the enzyme tyrosinase were formed in the structure of free-standing molecularly imprinted polymer membranes. Molecularly imprinted polymer membranes with the catalytic activity were obtained by co-polymerization of the complex Cu (II)–catechol–urocanic acid ethyl ester with (tri)ethyleneglycoldimethacrylate, and oligourethaneacrylate. Addition of the elastic component oligourethaneacrylate provided formation of the highly cross-linked polymer with the catalytic activity in a form of thin, flexible, and mechanically stable membrane. High accessibility of the artificial catalytic sites for the interaction with the analyzed phenol molecules was achieved due to addition of linear polymer (polyethyleneglycol Mw 20,000) to the initial monomer mixture before the polymerization. As a result, typical semi-interpenetrating polymer networks (semi-IPNs) were formed. The cross-linked component of the semi-IPN was represented by the highly cross-linked catalytic molecularly imprinted polymer, while the linear one was represented by polyethyleneglycol Mw 20,000. Extraction of the linear polymer from the fully formed semi-IPN resulted in formation of large pores in the membranes’ structure. Concentration of phenols in the analyzed samples was detected using universal portable device oxymeter with the oxygen electrode in a close contact with the catalytic molecularly imprinted polymer membrane as a transducer. The detection limit of phenols detection using the developed sensor system based on polymers–biomimics with the optimized composition comprised 0.063 mM, while the linear range of the sensor comprised 0.063–1 mM. The working characteristics of the portable sensor devices were investigated. Storage stability of sensor systems at room temperature comprised 12 months (87%). As compared to traditional methods of phenols detection the developed sensor system is characterized by simplicity of operation, compactness, an

Molecularly Imprinted Polymers for Cell Recognition

Since their conception 50 years ago, molecularly imprinted polymers (MIPs) have seen extensive development both in terms of synthetic routes and applications. Cells are perhaps the most challenging target for molecular imprinting. Although early work was based almost entirely around microprinting methods, recent developments have shifted towards epitope imprinting to generate MIP nanoparticles (NPs). Simultaneously, the development of techniques such as solid phase MIP synthesis has solved many historic issues of MIP production. This review briefly describes various approaches used in cell imprinting with a focus on applications of the created materials in imaging, drug delivery, diagnostics, and tissue engineering

Development of electrochemical sensors for the detection of photosystem inhibiting herbicides

The objective of the present work is the development of an amperometric sensor for detection of hydrogen peroxide and its integration with spinach chloroplasts for the further development of a sensor for herbicides. The design of the sensor employed screen- printing electrodes which are easily produced at the facilities available in Cranfield University. The hydrogen peroxide sensor has been based on horseradish peroxidase (HRP) as the catalytic element and hydroquinone as the mediator. HRP has been immobilised onto the sensor surface using a newly developed thioacetale-based polymer capable of covalent immobilisation of primary amines. A new HRP-based biosensor was screen-printed using a carbon/polymer mixture. Hydrogen peroxide concentrations were analysed at the reduction potential of hydroquinone (–0.3 V). The biosensors developed in this work had low detection limit of HB2BOB2B (0.1 µM), long term stability (they can be stored for 2 months at 4 P 0 PC) and good reproducibility of measurements (RSD ~ 5%). The hydrogen peroxide sensor has been further integrated with spinach chloroplasts in an attempt to create a sensor for photosynthesis-inhibiting herbicides. It was found however that the quantity of HB2BOB2B generated by chloroplasts in our experimental conditions was not sufficient to allow quantitative analysis. Due to this we have developed an alternative approach based on the electrochemistry of the Hill reaction. In this reaction the photosynthetic process and electron flow passing through photosystem II (PSII) is monitored through the quantity of reduced artificial electron acceptor. Upon illumination of the chloroplasts a signal from a reduced acceptor or mediator was recorded chronoamperometrically. The added herbicide inhibits the photosynthetic process and decreases the reduction of mediator. The decrease in measured current which is proportional to herbicide concentrations have been used for herbicide detection. Three mediators of Hill reaction were tested including 2,6 dichlorophenolindophenol (DCPIP) duroquinone and potassium ferricyanide. The optimal results were obtained using DCPIP. The optimal wavelength for the excitation of chloroplasts was 650 nm. The chloroplasts have been immobilised onto the sensor surface using cross-linking with glutaraldehyde and bovine serum albumin. The developed system allowed reliable detection of herbicides (RSD = 10%) with a detection limit of 1-8 nM depending on the type of herbicide. The sensor can be stored for 3 months at -80 P 0 PC. Preliminary measurements of river water samples using this sensor were also performed indicating good correlation between the data obtained with GC-MS and the chloroplast-based biosensor developed in this study.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Attenuation of quorum sensing using computationally designed polymers

It is generally accepted that the majority of Gram-negative and Gram-positive bacteria communicate via production and sensing of small signal molecules, autoinducers. The ability of bacteria to sense their population density is termed quorum sensing (QS). Quorum sensing controls certain phenotypic traits, particularly virulence factors and biofilm formation. In this project a new solution for the attenuation of quorum sensing which involves selective sequestering of the signal molecules using rationally designed synthetic polymers was explored.EThOS - Electronic Theses Online ServiceGBUnited Kingdo