Thesis

The single step purification and immobilisation of proteins on molecularly imprinted polymer supports

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Awarding institution
  • University of Strathclyde
Date of award
  • 2010
Thesis identifier
  • T12753
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Department, School or Faculty
Abstract
  • The molecular imprinting of proteins is a highly appealing area of research since polymers that can bind selectively to proteins can be applied in a number of interesting fields, including affinity separation, chemical sensing and molecular diagnostics. Generally speaking, proteins are significantly more difficult to imprint than low molar mass templates due to their physical size, thermal instability and their tendency to denature in the solvents used typically in molecular imprinting protocols. However, this has not deterred research in this area. Previous protein imprinting reports include the imprinting of gels, the surface imprinting of silica, and the epitope approach. In the Strathclyde approach to protein imprinting, protein in the form of an insoluble protein-coated microcrystal (PCMC) is used as the template. This PCMC strategy addresses many of the difficulties which can arise during the imprinting of proteins, including the thermal instability of proteins and their tendency to denature in many chemical environments. This novel synthetic approach has allowed for different types of protein, including enzymes, to be imprinted successfully. Molecularly imprinted solid-phase extraction (MISPE) has been used to demonstrate that MIPs prepared in this way can extract templated protein from an aqueous environment in an efficient manner, and also to probe the cross-selectivity of the imprinted sorbents for structurally-related and structurally non-related proteins. For imprinted enzymes, where catalytic activity is an important metric, once the enzyme has been bound to the imprinted polymer, the activity of the immobilised enzyme has been assessed and its stability monitored over various time and temperature ranges. It has been found that the enzyme remains catalytically active when bound to the imprinted polymer and indeed shows enhanced stability compared to the enzyme immobilised on non-imprinted polymers. Finally the selective extraction of enzyme from a complex biological sample (a cell lysate) has been demonstrated.
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Note
  • This thesis was previously held under moratorium from 4th May 2011 until 4th May 2013.
DOI
Date Created
  • 2010
Former identifier
  • 821486

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