title: Pluronic-lysozyme conjugates as anti-adhesive and antibacterial bifunctional polymers for surface coating.
abstract: This paper describes the preparation and characterization of polymer-protein conjugates composed of a synthetic triblock copolymer with a central polypropylene oxide (PPO) block and two terminal polyethylene oxide (PEO) segments, Pluronic F-127, and the antibacterial enzyme lysozyme attached to the telechelic groups of the PEO chains. Covalent conjugation of lysozyme proceeded via reductive amination of aldehyde functionalized PEO blocks (CHO-Pluronic) and the amine groups of the lysine residues in the protein. SDS-PAGE gel electrophoresis together with MALDI-TOF mass spectrometry analysis revealed formation of conjugates of one or two lysozyme molecules per Pluronic polymer chain. The conjugated lysozyme showed antibacterial activity towards Bacillus subtilis. Analysis with a quartz crystal microbalance with dissipation revealed that Pluronic-lysozyme conjugates adsorb in a brush conformation on a hydrophobic gold-coated quartz surface. X-ray photoelectron spectroscopy indicated surface coverage of 32% by lysozyme when adsorbed from a mixture of unconjugated Pluronic and Pluronic-lysozyme conjugate (ratio 99:1) and of 47% after adsorption of 100% Pluronic-lysozyme conjugates. Thus, bifunctional brushes were created, possessing both anti-adhesive activity due to the polymer brush, combined with the antibacterial activity of lysozyme. The coating having a lower degree of lysozyme coverage proved to be more bactericidal.
authors.affiliation: Department of Biomedical Engineering, W. J. Kolff Institute, FB40, University Medical Center Groningen and University of Groningen, P.O. Box 196, 9700 AD Groningen, The Netherlands.
authors.forename: Agnieszka K
authors.initials: AK
authors.lastname: Muszanska
authors.forename: Henk J
authors.initials: HJ
authors.lastname: Busscher
authors.forename: Andreas
authors.initials: A
authors.lastname: Herrmann
authors.forename: Henny C
authors.initials: HC
authors.lastname: van der Mei
authors.forename: Willem
authors.initials: W
authors.lastname: Norde
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000494:pharmacology; D001412:Bacillus subtilis / Q000187:drug effects; D009113:Muramidase / Q000737:chemistry* / Q000494:pharmacology; D056951:Photoelectron Spectroscopy; D020442:Poloxamer / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000900:Anti-Bacterial Agents; D011108:Polymers; D020442:Poloxamer; D009113:Muramidase