A series of polymer-bound antimicrobial peptides was prepared, and the peptides were tested for their antimicrobial activities. The immobilized peptides were prepared by a strategy that used solid-phase peptide synthesis that linked the carboxy-terminal amino acid with an ethylenediamine-modified polyamide resin (PepsynK). The acid-stable, permanent amide bond between the support and the nascent peptide renders the peptide resistant to cleavage from the support during the final acid-catalyzed deprotection step in the synthesis. Select immobilized peptides containing amino acid sequences that ranged from the naturally occurring magainin to simpler synthetic sequences with idealized secondary structures were excellent antimicrobial agents against several organisms. The immobilized peptides typically reduced the number of viable cells by Ն5 log units. We show that the reduction in cell numbers cannot be explained by the action of a soluble component. We observed no leached or hydrolyzed peptide from the resin, nor did we observe any antimicrobial activity in soluble extracts from the immobilized peptide. The immobilized peptides were washed and reused for repeated microbial contact and killing. These results suggest that the surface actions by magainins and structurally related antimicrobial peptides are sufficient for their lethal activities.Microbial contamination and possible infection of the host are major concerns in the area of therapeutic medical devices (13,15). Ongoing studies are directed toward understanding the mechanism of microbial adsorption and proliferation on material surfaces (13,15,40) and devising new approaches that can be used to avoid this microbial contamination (3,8,9,34). Sequestering the antimicrobial agent close to the device surface has been shown to be an effective approach to reducing the potential for catheter-related infections in patients (18). We believe that the immobilization of the antimicrobial agent by covalent bonding has the advantage of long-term stability and lower toxicity than that by a leach-or a release-based system (10, 36). We chose to explore whether a variety of peptide sequences ranging from naturally occurring magainin to synthetic idealized amphiphilic peptides, as described by DeGrado (7) and Blondelle and Houghton (1), and several closely related analogs would be bactericidal when they are bound to an insoluble support. Antimicrobial peptides such as the magainins, cecropins, and defensins are attractive candidates for such a support-bound antimicrobial agent because of their broad-spectrum activities, their relatively low cytotoxicities, and a body of basic evidence that suggests that the membrane surface is their lethal site of action (2,12,16,22,23,35,(37)(38)(39)41).In addition to seeking an improved approach to lower microbial proliferation on polymer surfaces, we were also interested in exploring an alternative experimental design for evaluating the mechanisms of action of these particular antimicrobial peptides. The stable chemical immobilization of t...