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    Pluronic–lysozyme conjugates as anti-adhesive and antibacterial bifunctional polymers for surface coating

    Year: 2011

    Journal: Biomaterials, Volume 32, Issue 26, September 2011, Pages 6333-6341, 20110906

    Authors: Muszanska A.K. 1, Busscher H.J. 1, Herrmann A. 2, van der Mei H.C. 1, Norde W. 1 3

    Last authors: Willem Norde

    Organizations: 1 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 2 Zernike Institute for Advanced Materials, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands 3 Laboratory of Physical Chemistry and Colloid Science, Wageningen University and Research Center, P.O. Box 8038, 6700 EK Wageningen, The Netherlands

    Country: The Netherlands, Holland

    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.