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Polymer-Mediated Cryopreservation of Bacteriophages
[Image: see text] 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 profile...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8672357/ https://www.ncbi.nlm.nih.gov/pubmed/34846863 http://dx.doi.org/10.1021/acs.biomac.1c01187 |
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author | Marton, Huba L. Styles, Kathryn M. Kilbride, Peter Sagona, Antonia P. Gibson, Matthew I. |
author_facet | Marton, Huba L. Styles, Kathryn M. Kilbride, Peter Sagona, Antonia P. Gibson, Matthew I. |
author_sort | Marton, Huba L. |
collection | PubMed |
description | [Image: see text] 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. |
format | Online Article Text |
id | pubmed-8672357 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-86723572021-12-15 Polymer-Mediated Cryopreservation of Bacteriophages Marton, Huba L. Styles, Kathryn M. Kilbride, Peter Sagona, Antonia P. Gibson, Matthew I. Biomacromolecules [Image: see text] 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. American Chemical Society 2021-11-30 2021-12-13 /pmc/articles/PMC8672357/ /pubmed/34846863 http://dx.doi.org/10.1021/acs.biomac.1c01187 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Marton, Huba L. Styles, Kathryn M. Kilbride, Peter Sagona, Antonia P. Gibson, Matthew I. Polymer-Mediated Cryopreservation of Bacteriophages |
title | Polymer-Mediated Cryopreservation of Bacteriophages |
title_full | Polymer-Mediated Cryopreservation of Bacteriophages |
title_fullStr | Polymer-Mediated Cryopreservation of Bacteriophages |
title_full_unstemmed | Polymer-Mediated Cryopreservation of Bacteriophages |
title_short | Polymer-Mediated Cryopreservation of Bacteriophages |
title_sort | polymer-mediated cryopreservation of bacteriophages |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8672357/ https://www.ncbi.nlm.nih.gov/pubmed/34846863 http://dx.doi.org/10.1021/acs.biomac.1c01187 |
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