Investigation into scalable and efficient enterotoxigenic Escherichia coli bacteriophage production

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 phag...

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Bibliographic Details
Published in:	Scientific reports Vol. 14; no. 1; p. 3618
Main Authors:	Wiebe, Katie G., Cook, Bradley W. M., Lightly, Tasia J., Court, Deborah A., Theriault, Steven S.
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 13-02-2024 Nature Publishing Group Nature Portfolio
Subjects:	631/326/1321 631/326/596 631/61/252 Antibiotic resistance Bacteria Bacteriophages Bioreactors Cell culture E coli Enterotoxigenic Escherichia coli Escherichia coli Humanities and Social Sciences Lysis multidisciplinary Multiplicity of infection Optical density Phages Science Science (multidisciplinary) Temperature
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	2045-2322 2045-2322
DOI:	10.1038/s41598-024-53276-w