Abstract
It has been over 100 years since bacteriophages (phages) were used as a human therapeutic. Since then, phage production has dramatically evolved. Current phage preparations have fewer adverse effects due to their low bacterial toxin content. As a result, therapeutic phages have become a predominant class of new antimicrobials and are being widely used for compassionate treatment of multidrug-resistant (MDR) infections. We describe herein a protocol for the production and ultrapurification of phages. By this technique, it is possible for a lab experienced with the process to produce >109 plaque-forming units (PFU) per mL of Gram-negative phages that meet FDA endotoxins limits for intravenous infusions in as little as 48 hours. We provide illustrations of the process and tips on how to safely remove bacterial toxins from phage lysates. Although dependent on the phage strain, the approach described can rapidly generate and purify phages for a variety of applications.
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Acknowledgments
We would like to acknowledge Nino Grdzelishvili for her feedback and suggestions. TL is a recipient of the ARCS Foundation Scholarship.
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Luong, T., Sue, A.D., Roach, D.R. (2024). Rapid Bench to Bedside Therapeutic Bacteriophage Production. In: Azeredo, J., Sillankorva, S. (eds) Bacteriophage Therapy. Methods in Molecular Biology, vol 2734. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3523-0_5
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DOI: https://doi.org/10.1007/978-1-0716-3523-0_5
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