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Polymer-mediated cryopreservation of bacteriophages

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Marton, Huba L., Styles, Kathryn, Kilbride, Peter, Sagona, Antonia P. and Gibson, Matthew I. (2021) Polymer-mediated cryopreservation of bacteriophages. Biomacromolecules, 22 (12). pp. 5281-5289. doi:10.1021/acs.biomac.1c01187

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Official URL: http://dx.doi.org/10.1021/acs.biomac.1c01187

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Abstract

Bacteriophages (phages, bacteria-specific viruses) have biotechnological and therapeutic potential. To apply phages as pure or heterogeneous mixtures, it is essential to have a robust mechanism for transport and storage, with different phages having very different stability profiles across storage conditions. For many biologics, cryopreservation is employed for long-term storage and cryoprotectants are essential to mitigate cold-induced damage. Here, we report that poly(ethylene glycol) can be used to protect phages from cold damage, functioning at just 10 mg·mL–1 (∼1 wt %) and outperforms glycerol in many cases, which is a currently used cryoprotectant. Protection is afforded at both −20 and −80 °C, the two most common temperatures for frozen storage in laboratory settings. Crucially, the concentration of the polymer required leads to frozen solutions at −20 °C, unlike 50% glycerol (which results in liquid solutions). Post-thaw recoveries close to 100% plaque-forming units were achieved even after 2 weeks of storage with this method and kill assays against their bacterial host confirmed the lytic function of the phages. Initial experiments with other hydrophilic polymers also showed cryoprotection, but at this stage, the exact mechanism of this protection cannot be concluded but does show that water-soluble polymers offer an alternative tool for phage storage. Ice recrystallization inhibiting polymers (poly(vinyl alcohol)) were found to provide no additional protection, in contrast to their ability to protect proteins and microorganisms which are damaged by recrystallization. PEG’s low cost, solubility, well-established low toxicity/immunogenicity, and that it is fit for human consumption at the concentrations used make it ideal to help translate new approaches for phage therapy.

Item Type: Journal Article
Subjects: Q Science > QD Chemistry
Q Science > QR Microbiology
T Technology > TP Chemical technology
Divisions: Faculty of Science > Chemistry
Faculty of Science > Life Sciences (2010- )
Library of Congress Subject Headings (LCSH): Bacteriophages , Microorganisms -- Cryopreservation, Polyethylene glycol, Water-soluble polymers
Journal or Publication Title: Biomacromolecules
Publisher: American Chemical Society
ISSN: 1525-7797
Official Date: 13 December 2021
Dates:
DateEvent
13 December 2021Published
30 November 2021Available
11 November 2021Accepted
Volume: 22
Number: 12
Page Range: pp. 5281-5289
DOI: 10.1021/acs.biomac.1c01187
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Open Access
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
BB/M01116X/1[BBSRC] Biotechnology and Biological Sciences Research Councilhttp://dx.doi.org/10.13039/501100000268
UNSPECIFIEDCytivahttps://www.cytivalifesciences.com/en/gb/about-us
866056European Research Councilhttp://dx.doi.org/10.13039/501100000781
191037Royal Societyhttp://dx.doi.org/10.13039/501100000288
EP/S001255/1[EPSRC] Engineering and Physical Sciences Research Councilhttp://dx.doi.org/10.13039/501100000266

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