Cell-free production of personalized therapeutic phages targeting multidrug-resistant bacteria

Emslander, Quirin; Vogele, Kilian; Braun, Peter; Stender, Jana; Willy, Christian; Joppich, Markus; Hammerl, Jens A.; Abele, Miriam; Meng, Chen; Pichlmair, Andreas; Ludwig, Christina; Bugert, Joachim J.; Simmel, Friedrich C. und Westmeyer, Gil G. (2022): Cell-free production of personalized therapeutic phages targeting multidrug-resistant bacteria. In: Cell Chemical Biology, Bd. 29, Nr. 9: S. 1434-1445

Abstract

Bacteriophages are potent therapeutics against biohazardous bacteria, which rapidly develop multidrug resistance. However, routine administration of phage therapy is hampered by a lack of rapid production, safe bioengineering, and detailed characterization of phages. Thus, we demonstrate a comprehensive cell-free platform for personalized production, transient engineering, and proteomic characterization of a broad spectrum of phages. Using mass spectrometry, we validated hypothetical and non-structural proteins and could also monitor the protein expression during phage assembly. Notably, a few microliters of a one-pot reaction produced effective doses of phages against enteroaggregative Escherichia coli (EAEC), Yersinia pestis, and Klebsiella pneumoniae. By co-expressing suitable host factors, we could extend the range of cell-free production to phages targeting gram-positive bacteria. We further introduce a non-genomic phage engineering method, which adds functionalities for only one replication cycle. In summary, we expect this cell-free methodology to foster reverse and forward phage engineering and customized production of clin-ical-grade bacteriophages.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Mathematik, Informatik und Statistik > Informatik
• Themengebiete: 000 Informatik, Informationswissenschaft, allgemeine Werke > 004 Informatik
• ISSN: 2451-9456
• Sprache: Englisch
• DFG: Gefördert durch die Deutsche Forschungsgemeinschaft (DFG) - 364653263