Investigation into scalable and efficient enterotoxigenic Escherichia coli bacteriophage production.

Autor: Wiebe KG; Cytophage Technologies Inc., Winnipeg, MB, Canada.; Department of Microbiology, University of Manitoba, Winnipeg, MB, Canada., Cook BWM; Cytophage Technologies Inc., Winnipeg, MB, Canada., Lightly TJ; Cytophage Technologies Inc., Winnipeg, MB, Canada., Court DA; Department of Microbiology, University of Manitoba, Winnipeg, MB, Canada., Theriault SS; Cytophage Technologies Inc., Winnipeg, MB, Canada. Steven@cytophage.com.; Department of Microbiology, University of Manitoba, Winnipeg, MB, Canada. Steven@cytophage.com.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2024 Feb 13; Vol. 14 (1), pp. 3618. Date of Electronic Publication: 2024 Feb 13.
DOI: 10.1038/s41598-024-53276-w
Abstrakt: As the demand for bacteriophage (phage) therapy increases due to antibiotic resistance in microbial pathogens, strategies and methods for increased efficiency, large-scale phage production need to be determined. To date, very little has been published on how to establish scalable production for phages, while achieving and maintaining a high titer in an economical manner. The present work outlines a phage production strategy using an enterotoxigenic Escherichia coli-targeting phage, 'Phage75', and accounts for the following variables: infection load, multiplicity of infection, temperature, media composition, harvest time, and host bacteria. To streamline this process, variables impacting phage propagation were screened through a high-throughput assay monitoring optical density at 600 nm (OD 600 ) to indirectly infer phage production from host cell lysis. Following screening, propagation conditions were translated in a scalable fashion in shake flasks at 0.01 L, 0.1 L, and 1 L. A final, proof-of-concept production was then carried out in a CellMaker bioreactor to represent practical application at an industrial level. Phage titers were obtained in the range of 9.5-10.1 log 10 PFU/mL with no significant difference between yields from shake flasks and CellMaker. Overall, this suggests that the methodology for scalable processing is reliable for translating into large-scale phage production.
(© 2024. The Author(s).)
Databáze: MEDLINE
Externí odkaz: