Dataset Information

A cell-free platform for characterizing and producing therapeutic bacteriophages against biohazardous and antibiotic-resistant bacteria

ABSTRACT: Bacteriophages are potent therapeutics against biohazardous bacteria that are rapidly acquiring multidrug resistance. However, routine administration of bacteriophage therapy is currently impeded by a lack of safe phage production methodologies and insufficient phage characterization. We thus developed a versatile cell-free platform for host-independent production of phages targeting gram-positive and gram-negative bacteria. A few microliters of a one-pot reaction produces effective doses of phages against potentially antibiotic-resistant bacteria such as enterohemorrhagic E. coli (EAEC) and Yersinia pestis, which also possibly pose threats as biological warfare agents. We also introduce a method for transient, non-genomic phage engineering to safely confer additional functions, such as a purification tag or bioluminescence for host detection, for only one replication cycle. Using high-resolution and time-resolved mass spectrometry, we validated the expression of 40 hypothetical proteins from two different phages (T7 and CLB-P3) and identified genes in the genome of phage T7 that express exceptionally late during phage replication. Our comprehensive methodology thus allows for accelerated reverse and forward phage engineering as well as for safe and customized production of clinical-grade therapeutic bacteriophages.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Escherichia Coli Enterobacteria Phage T7 Bacteria Eggerthella Phage P3

SUBMITTER: Christina Ludwig

LAB HEAD: Chen Meng

PROVIDER: PXD019543 | Pride | 2022-07-20

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	BBM_197_P117_04_MIA_001_R2.raw	Raw
	BBM_197_P117_04_MIA_002_R2.raw	Raw
	BBM_197_P117_04_MIA_003_R2.raw	Raw
	BBM_197_P117_04_MIA_004_R2.raw	Raw
	BBM_197_P117_04_MIA_005_R2.raw	Raw

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Publications

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

Emslander Quirin Q Vogele Kilian K Braun Peter P Stender Jana J Willy Christian C Joppich Markus M Hammerl Jens A JA Abele Miriam M Meng Chen C Pichlmair Andreas A Ludwig Christina C Bugert Joachim J JJ Simmel Friedrich C FC Westmeyer Gil G GG

Cell chemical biology 20220711 9

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-str ...[more]

PMID: 35820417

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