학술논문
Rapid assessment of changes in phage bioactivity using dynamic light scattering
Research Report
Research Report
Document Type
Academic Journal
Author
Dharmaraj, Tejas; Kratochvil, Michael J.; Pourtois, Julie D.; Chen, Qingquan; Hajfathalian, Maryam; Hargil, Aviv; Lin, Yung-Hao; Evans, Zoe; Oromi-Bosch, Agnes; Berry, Joel D.; McBride, Robert; Haddock, Naomi L.; Holman, Derek R.; van Belleghem, Jonas D.; Chang, Tony H.; Barr, Jeremy J.; Lavigne, Rob; Heilshorn, Sarah C.; Blankenberg, Francis G.; Bollyky, Paul L.
Source
PNAS Nexus. December 2023, Vol. 2 Issue 12
Subject
Language
English
ISSN
2752-6542
Abstract
Introduction As antimicrobial-resistant (AMR) pathogens have become increasingly prevalent (1-3), there is great interest in novel approaches to treat bacterial infections. Bacteriophages (phages), viruses that kill bacteria, offer a promising [...]
Extensive efforts are underway to develop bacteriophages as therapies against antibiotic-resistant bacteria. However, these efforts are confounded by the instability of phage preparations and a lack of suitable tools to assess active phage concentrations over time. In this study, we use dynamic light scattering (DLS) to measure changes in phage physical state in response to environmental factors and time, finding that phages tend to decay and form aggregates and that the degree of aggregation can be used to predict phage bioactivity. We then use DLS to optimize phage storage conditions for phages from human clinical trials, predict bioactivity in 50-y-old archival stocks, and evaluate phage samples for use in a phage therapy/wound infection model. We also provide a web application (Phage-Estimator of Lytic Function) to facilitate DLS studies of phages. We conclude that DLS provides a rapid, convenient, and nondestructive tool for quality control of phage preparations in academic and commercial settings. Keywords: bacteriophage therapy, dynamic light scattering, bacteriophage decay, aggregation
Extensive efforts are underway to develop bacteriophages as therapies against antibiotic-resistant bacteria. However, these efforts are confounded by the instability of phage preparations and a lack of suitable tools to assess active phage concentrations over time. In this study, we use dynamic light scattering (DLS) to measure changes in phage physical state in response to environmental factors and time, finding that phages tend to decay and form aggregates and that the degree of aggregation can be used to predict phage bioactivity. We then use DLS to optimize phage storage conditions for phages from human clinical trials, predict bioactivity in 50-y-old archival stocks, and evaluate phage samples for use in a phage therapy/wound infection model. We also provide a web application (Phage-Estimator of Lytic Function) to facilitate DLS studies of phages. We conclude that DLS provides a rapid, convenient, and nondestructive tool for quality control of phage preparations in academic and commercial settings. Keywords: bacteriophage therapy, dynamic light scattering, bacteriophage decay, aggregation