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Efficient synthetic receptors for endotoxin detection using molecular imprinting

Huckle, Danielle Louise 2019. Efficient synthetic receptors for endotoxin detection using molecular imprinting. PhD Thesis, Cardiff University.
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Abstract

Sepsis remains a poorly understood and potentially devastating medical syndrome. Unlike other medical emergencies there is no definitive, measureable biomarker. Lipopolysaccharide (LPS) is widely implicated in the pathogenesis of sepsis. It is not routinely measured owing to a lack of reliable diagnostic assay. Molecular imprinting is a novel approach to LPS detection. It describes the generation of synthetic, polymeric receptors through the polymerisation of monomers around a template molecule. The resultant polymers possess recognition properties akin to an antibody with the ability to recognise the original target molecule. LPS is not a suitable template for conventional molecular imprinting techniques, therefore, techniques that circumvent problems associated with the imprinting of biological macromolecules are needed. Modified approaches that localise the template at a surface via chemical immobilisation at a solid-solvent interface (surface imprinting on a solid support) or via self-assembly at a solvent-solvent interface (microfluidic techniques plus microwave-assisted polymerisation) have been investigated. Polymyxin B (PMB), a peptide antibiotic, has high affinity for LPS and was utilised in this study to help localise LPS at an interface under a variety of experimental conditions; including the synthesis of a polymerisable PMB (p-PMB). A synthesised p-PMB polymer was subsequently found able to bind LPS from solution with an apparent Kd of 0.327 μM. Furthermore, using a bespoke microfluidic/microwave hybrid system, with benzethonium chloride (BC) used as a LPS surrogate, BC imprinted polymer beads were produced. Subsequent binding experiments demonstrated ability of the imprinted bead to bind to the original template with a Bmax of 0.014 nmol/mg and an apparent Kd of 0.536 μM.

Item Type: Thesis (PhD)
Date Type: Completion
Status: Unpublished
Schools: Medicine
Date of First Compliant Deposit: 10 September 2019
Last Modified: 10 Sep 2019 09:46
URI: http://orca-mwe.cf.ac.uk/id/eprint/125363

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