title: Immobilization of Active Bacteriophages on Polyhydroxyalkanoate Surfaces.
abstract: A rapid, efficient technique for the attachment of bacteriophages (phages) onto polyhydroxyalkanoate (PHA) surfaces has been developed and compared to three reported methods for phage immobilization. Polymer surfaces were modified to facilitate phage attachment using (1) plasma treatment alone, (2) plasma treatment followed by activation by 1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (sulfo-NHS), (3) plasma-initiated acrylic acid grafting, or (4) plasma-initiated acrylic acid grafting with activation by EDC and sulfo-NHS. The impact of each method on the surface chemistry of PHA was investigated using contact angle analysis and X-ray photoelectron spectroscopy. Each of the four treatments was shown to result in both increased hydrophilicity and in the modification of the surface functional groups. Modified surfaces were immersed in suspensions of phage T4 for immobilization. The highest level of phage binding was observed for the surfaces modified by plasma treatment alone. The change in chemical bond states observed for surfaces that underwent plasma treatment is suspected to be the cause of the increased binding of active phages. Plasma-treated surfaces were further analyzed through phage-staining and fluorescence microscopy to assess the surface density of immobilized phages and their capacity to capture hosts. The infective capability of attached phages was confirmed by exposing the phage-immobilized surfaces to the host bacteria Escherichia coli in both plaque and infection dynamic assays. Plasma-treated surfaces with immobilized phages displayed higher infectivity than surfaces treated with other methods; in fact, the equivalent initial multiplicity of infection was 2 orders of magnitude greater than with other methods. Control samples - prepared by immersing polymer surfaces in phage suspensions (without prior plasma treatment) - did not show any bacterial growth inhibition, suggesting they did not bind phages from the suspension.
authors.affiliation: Department of Chemical and Material Engineering, University of Alberta , 9211 116 Street NW, Edmonton, Alberta T6G 1H9, Canada.
authors.forename: Chanchan
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Chemical and Material Engineering, University of Alberta , 9211 116 Street NW, Edmonton, Alberta T6G 1H9, Canada.
authors.forename: Dominic
authors.initials: D
authors.lastname: Sauvageau
authors.affiliation: Department of Chemical and Material Engineering, University of Alberta , 9211 116 Street NW, Edmonton, Alberta T6G 1H9, Canada.
authors.forename: Anastasia
authors.initials: A
authors.lastname: Elias
mesh_terms: D017122:Bacteriophage T4 / Q000737:chemistry* / Q000378:metabolism; D001435:Bacteriophages / Q000737:chemistry*; D015374:Biosensing Techniques; D004926:Escherichia coli / Q000737:chemistry; D056951:Photoelectron Spectroscopy; D054813:Polyhydroxyalkanoates / Q000737:chemistry*; D013388:Succinimides / Q000737:chemistry; D013499:Surface Properties
chemical_list: D054813:Polyhydroxyalkanoates; D013388:Succinimides; C035761:N-hydroxysulfosuccinimide
keywords: active material; bacteriophage immobilization; detection of bacteria; inhibition of E. coli; phage binding; surface modification