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

2011; Elsevier BV; Volume: 32; Issue: 26 Linguagem: Inglês

10.1016/j.biomaterials.2011.05.016

1878-5905

Agnieszka K. Muszanska, Henk J. Busscher, Andreas Herrmann, Henny C. van der Mei, Willem Norde,

Antimicrobial Peptides and Activities

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.