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Mechanisms for covalent immobilization of horseradish peroxi-dase on ion beam treated polyethylene | | Alexey V. Kondyurin
; Pourandokht Naseri
; Jennifer M. R. Tilley
; Neil J. Nosworthy
; Marcela M.M. Bilek
; David R. McKenzie
; | | Date: |
14 Oct 2011 | | Abstract: | The mechanism that provides the observed strong binding of biomolecules to
polymer sur-faces modified by ion beams is investigated. The surface of
polyethylene (PE) was modified by plasma immersion ion implantation with
nitrogen ions. Structure changes including car-bonization and oxidation were
observed in the modified surface layer of PE by Raman spec-troscopy, FTIR ATR
spectroscopy, atomic force microscopy, surface energy measurement and XPS
spectroscopy. An observed high surface energy of the modified polyethylene was
attributed to the presence of free radicals on the surface. The surface energy
decay with stor-age time after PIII treatment was explained by a decay of the
free radical concentration while the concentration of oxygen-containing groups
increased with storage time. Horseradish per-oxidase was covalently attached
onto the modified PE surface. The enzymatic activity of co-valently attached
protein remained high. A mechanism based on the covalent attachment by the
reaction of protein with free radicals in the modified surface is proposed.
Appropriate blocking agents can block this reaction. All aminoacid residues can
take part in the covalent attachment process, providing a universal mechanism
of attachment for all proteins. The long-term activity of the modified layer to
attach protein (at least 2 years) is explained by stabilisa-tion of unpaired
electrons in sp2 carbon structures. The native conformation of attached
pro-tein is retained due to hydrophilic interactions in the interface region. A
high concentration of free radicals on the surface can give multiple covalent
bonds to the protein molecule and de-stroy the native conformation and with it
the catalytic activity. The universal mechanism of protein attachment to free
radicals could be extended to various methods of radiation damage of polymers. | | Source: | arXiv, 1110.3125 | | Services: | Forum | Review | PDF | Favorites | |
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