Design and Characterization of All-In-One Artificial Protein Platform Toward Novel Antimicrobial Materials

Abstract

Antimicrobial resistance is a crisis affecting millions of people each year around the globe and is progressively getting worse, but there are too few promising solutions in the pipeline. Antimicrobial peptides (AMPs) are an exciting alternative to antibiotics due to their broad-spectrum activity, but in vivo instability, such as proteolysis, has largely prevented their clinical approval. AMP drug delivery systems have made gains in stabilizing AMPs, but their complexity and high expense have been barriers to large-scale clinical translation. Here, we present an all-in-one artificial protein platform that simplified synthesis and processing and has the functionality to form micelle structures and hydrogels. We utilize genetic engineering and artificial proteins to form a single polymer strand that contains the material forming component, spacer component, and antimicrobial peptide such that conjugation steps are unnecessary. The use of elastin-like polypeptides as the representative artificial protein exploits an inexpensive protein purification strategy that avoids expensive chromatography. We showed that the all-in-one artificial protein platform can target the mechanical properties of natural tissues by manipulating the crosslinking formulations, we evaluated the biocompatibility of the material against fibroblasts, and we demonstrated the protein can form micelles at body temperature. Furthermore, we devised a method to improve the elastic moduli of hydrogels when self-assembled structures disrupted the efficiency of crosslinking. Altogether, these data provide the foundation toward utilizing the all-in-one artificial protein platform to stabilize AMPs and enhance AMP efficacy for clinical applications, such as tissue engineering and drug delivery.