We report the development of an automated chemical reactor for solid-phase synthesis of MIP NPs in water. Operational parameters are under computer control, requiring minimal operator intervention. In this study, "ready for use" MIP NPs with sub-nanomolar affinity are prepared against pepsin A, trypsin and α-amylase in only 4 hours.The extent of research into biomedical and diagnostic nanotechnology is impressive, 1 despite currently very few examples of biomedical nanomaterials have been approved by the FDA. 2 Apart from the potential toxicity, the usage of nanomaterials in therapy and diagnostics finds a strong opponent in the well-established leadership of "safer" natural antibodies. 3 Industrial manufacturing of antibodies, however, is logistically complex and expensive, especially for applications in therapy. 4 Potential immunogenic adverse reactions, low stability and poor performance in non-physiological conditions restrict the application of antibodies to specific niche areas and shorten their shelf-life. 5,6 Finally, natural antibodies can be difficult to integrate with assays and sensors. 6,7 A possible alternative are MIP NPs, which share key characteristics with antibodies such as aqueous solubility, size, affinity and selectivity for the target. Additionally, solutions of MIP NPs can be handled similarly to solutions of antibodies, with the advantage of stability and robustness. 8 Moreover, operational parameters for producing MIP NPs can be carefully controlled. 9 There is, however, lack of a generic protocol for the synthesis of MIP NPs with specific characteristics, 8 which restricts their practical applications, despite recent reports of biological activity 10 and in vivo detoxification 11 hint at a great potential of MIP NPs in the nanomedicine arena.With the aim to provide a method that is scalable, reproducible and controlled, we have focused our efforts on the development of synthetic strategies for producing highperformance MIP NPs. The most suitable protocol should in our view include affinity purification with immobilised template to remove monomers/low-affinity NPs from the product. 12 Even better results would be obtained if the immobilised template is used also in polymerisation, as this will ensure high-binding site affinity and prevent the contamination 13 This approach would also add advantages of easy automation and the potential for scaling-up production.Recently we have reported the first successful example of solid-phase synthesis of MIP NPs using living-polymerisation chemistry, performed in organic solvent under UV irradiationconditions which are favourable for the imprinting of small molecules. 14 Imprinting of highmolecular weight targets such as proteins, polysaccharides and DNA, however, would require an aqueous environment to preserve their structure during polymerisation. 15Here we report the development of a reactor for MIP NPs preparation using chemical polymerisation performed in specifically mild aqueous conditions (monomer concentration: 6.5 mM) for p...
The synthesis of core-shell molecularly imprinted polymer nanoparticles (MIP NPs) has been performed using a novel solid-phase approach on immobilised templates. The same solid phase also acts as protective functionality for high affinity binding sites during subsequent derivatisation/shell formation. This procedure allows for the rapid synthesis, controlled separation and purification of high-affinity materials, with each production cycle taking just 2 hours. The aim of this approach is to synthesise uniformly-sized imprinted materials at the nanoscale which can be readily grafted with various polymers without affecting their affinity and specificity. For demonstration purposes we grafted anti-melamine MIP NPs with coatings which introduce the following surface characteristics: high polarity (PEG methacrylate); electro-activity (vinyl ferrocene); fluorescence (eosin acrylate); thiol groups (pentaerythritol tetrakis(3-mercaptopropionate)). The method has broad applicability and can be used to produce multifunctional imprinted nanoparticles with potential for further application in the biosensors, diagnostics and biomedical fields and as an alternative to natural receptors.
An optical fibre long period grating (LPG) sensor modified with molecularly imprinted polymer nanoparticles (nanoMIPs) for the specific detection of antibiotics is presented. The operation of the sensor is based on the measurement of changes in refractive index induced by the interaction of nanoMIPs deposited onto the cladding of the LPG with free vancomycin (VA). The binding of nanoMIPs to vancomycin was characterised by a binding constant of 4.3 ± 0.1 × 10(-8) M. The lowest concentration of analyte measured by the fibre sensor was 10 nM. In addition, the sensor exhibited selectivity, as much smaller responses were obtained for high concentrations (∼700 μM) of other commonly prescribed antibiotics such as amoxicillin, bleomycin and gentamicin. In addition, the response of the sensor was characterised in a complex matrix, porcine plasma, spiked with 10 μM of VA.
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