Abstract
Glycidyl methacrylate (GMA) emulsion-templated porous polymers (polyHIPEs) were prepared by thermal and photopolymerisation and derivatised with morpholine, tris(2-aminoethylamine) and a bisamino-PEG homobifunctional molecule. The extent of the functionalization reactions was investigated by a range of qualitative and quantitative techniques (FTIR, CHN analysis, titration, XPS, HR-MAS NMR spectroscopy, ninhydrin assay and Fmoc number determination) and was found to be excellent for small molecule amines (up to 89% conversion) but low for the reaction with PEG (2% conversion). This was ascribed to the high exclusion volume of the PEG chains in solution. Proteinase K (Pro K) was subsequently immobilized covalently onto the GMA polyHIPE material, both directly via reaction with surface epoxy groups and indirectly by activation of the pendent amine groups of PEGylated polyHIPE with glutaraldehyde then reductive amination with the enzyme. The activity of the supported enzymes was determined by a continuous electrochemical assay involving the hydrolysis of N-acetyl-l-tyrosine ethyl ester. The directly immobilized Pro K was found to have an activity of only 3.6 U/g whereas the activity of the enzyme immobilized via the PEG linker was much higher (up to 78 U/g).
Highlights
Functional polymers can be prepared by the homopolymerization, copolymerization or grafting of a reactive monomer [1e3]
Having established that glycidyl methacrylate (GMA)-based polyHIPEs can be reliably functionalised with nucleophilic amines, we explored the possibility of attaching a homobifunctional bisamino-PEG linker (O,O’-bis(3-aminopropyl)polyethylene glycol) with which to anchor enzymes
Photopolymerised GMA-containing polyHIPE materials have been post-functionalized with a range of amine nucleophiles
Summary
Functional polymers can be prepared by the homopolymerization, copolymerization or grafting of a reactive monomer [1e3]. Poly(glycidyl methacrylate) (GMA) is an important functional polymer due to the ability of its epoxy group to react with a range of nucleophiles [1]. This has led to the preparation of porous GMA polymers for use as bioreactors and for protein separation [4e6]. Materials have been prepared by thermal or photochemical polymerisation of GMA as well as the grafting of GMA from a polyHIPE surface [16e21] These materials have been observed to be capable of functionalization with nucleophiles and have been used for protein separation [18,22]. The activity of the immobilized Pro K was monitored with a continuous electrochemical assay, monitoring the hydrolysis of N-acetyl-L-tyrosine ethyl ester monohydrate
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