Rapid assessment of changes in phage bioactivity using dynamic light scattering

Author:

Dharmaraj Tejas12ORCID,Kratochvil Michael J3ORCID,Pourtois Julie D4ORCID,Chen Qingquan1,Hajfathalian Maryam1,Hargil Aviv1,Lin Yung-Hao5ORCID,Evans Zoe1ORCID,Oromí-Bosch Agnès6,Berry Joel D6,McBride Robert6,Haddock Naomi L1,Holman Derek R7ORCID,van Belleghem Jonas D1ORCID,Chang Tony H1ORCID,Barr Jeremy J8ORCID,Lavigne Rob9ORCID,Heilshorn Sarah C3ORCID,Blankenberg Francis G10,Bollyky Paul L1ORCID

Affiliation:

1. Division of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Beckman Center for Molecular and Genetic Medicine , Stanford, CA 94305 , USA

2. Sarafan ChEM-H, Stanford University , Stanford, CA 94305 , USA

3. Department of Materials Science and Engineering, Stanford University , Stanford, CA 94305 , USA

4. Department of Biology, Hopkins Marine Station, Stanford University , Pacific Grove, CA 93950 , USA

5. Department of Chemical Engineering, Stanford University , Stanford, CA 94305 , USA

6. Felix Biotechnology , South SanFrancisco, CA 94080 , USA

7. Division of Gastroenterology and Hepatology, Department of Medicine, Stanford University School of Medicine , Stanford, CA 94305 , USA

8. School of Biological Sciences, Monash University , Clayton, VIC 3800 , Australia

9. Department of Biosystems, KU Leuven , Leuven 3001 , Belgium

10. Division of Pediatric Radiology and Nuclear Medicine, Department of Radiology, Lucile Packard Children's Hospital , Stanford, CA 94305 , USA

Abstract

Abstract 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.

Funder

National Institutes of Health

Cystic Fibrosis Foundation

Emerson Collective

Stanford University Medical Scientist Training Program

Stanford Interdisciplinary Graduate Fellowship

Gold Family Graduate Fellow

Publisher

Oxford University Press (OUP)

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