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

Autor:	Emslander Q; Institute of Virology, TUM School of Medicine, Technical University of Munich, 81675 Munich, Germany., Vogele K; Department of Physics, Technical University of Munich, 85748 Garching, Germany., Braun P; Bundeswehr Institute of Microbiology, 80937 Munich, Germany., Stender J; Bundeswehr Institute of Microbiology, 80937 Munich, Germany., Willy C; Bundeswehrkrankenhaus Berlin, 10115 Berlin, Germany., Joppich M; Department of Informatics, Ludwigs-Maximilian-Universität München, Amalienstraße 17, 80333 Munich, Germany., Hammerl JA; Department of Biological Safety, German Federal Institute for Risk Assessment, Max-Dohrn Str. 8-10, D-10589 Berlin, Germany., Abele M; BayBioMS, Bavarian Center for Biomolecular Mass Spectrometry, Technical University of Munich, 85354 Freising, Germany., Meng C; BayBioMS, Bavarian Center for Biomolecular Mass Spectrometry, Technical University of Munich, 85354 Freising, Germany., Pichlmair A; Institute of Virology, TUM School of Medicine, Technical University of Munich, 81675 Munich, Germany., Ludwig C; BayBioMS, Bavarian Center for Biomolecular Mass Spectrometry, Technical University of Munich, 85354 Freising, Germany., Bugert JJ; Bundeswehr Institute of Microbiology, 80937 Munich, Germany., Simmel FC; Department of Physics, Technical University of Munich, 85748 Garching, Germany., Westmeyer GG; Department of Chemistry & TUM School of Medicine, Technical University of Munich, 85748 Garching, Germany; Institute for Synthetic Biomedicine, Helmholtz Zentrum München, 85764 Oberschleißheim, Germany. Electronic address: gil.westmeyer@tum.de.
Jazyk:	angličtina
Zdroj:	Cell chemical biology [Cell Chem Biol] 2022 Sep 15; Vol. 29 (9), pp. 1434-1445.e7. Date of Electronic Publication: 2022 Jul 11.
DOI:	10.1016/j.chembiol.2022.06.003
Abstrakt:	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 clinical-grade bacteriophages. Competing Interests: Declaration of interests A patent application has been filed by the authors’ employer, the Technical University of Munich on behalf of Q.E., K.V., and F.C.S.. K.V. is currently leading the Invitris team funded by an EXIST Transfer for Research grant by the Federal Ministry of Economy and Climate Action to pursue work related to this article. Q.E. is a scientific advisor to the team and F.C.S., and G.G.W formally serve as mentors at TUM. (Copyright © 2022 Elsevier Ltd. All rights reserved.)
Databáze:	MEDLINE
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