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使用动态光散射快速评估噬菌体生物活性的变化

Rapid assessment of changes in phage bioactivity using dynamic light scattering.

作者信息

Dharmaraj Tejas, Kratochvil Michael J, Pourtois Julie D, Chen Qingquan, Hajfathalian Maryam, Hargil Aviv, Lin Yung-Hao, Evans Zoe, Oromí-Bosch Agnès, 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

机构信息

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.

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

出版信息

PNAS Nexus. 2023 Nov 27;2(12):pgad406. doi: 10.1093/pnasnexus/pgad406. eCollection 2023 Dec.

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.

摘要

人们正在付出巨大努力来开发噬菌体作为对抗抗生素耐药细菌的疗法。然而,这些努力因噬菌体制剂的不稳定性以及缺乏合适的工具来长期评估活性噬菌体浓度而受到困扰。在本研究中,我们使用动态光散射(DLS)来测量噬菌体物理状态随环境因素和时间的变化,发现噬菌体倾向于衰变并形成聚集体,并且聚集程度可用于预测噬菌体生物活性。然后,我们使用DLS来优化来自人体临床试验的噬菌体的储存条件,预测50年历史存档样本中的生物活性,并评估用于噬菌体疗法/伤口感染模型的噬菌体样本。我们还提供了一个网络应用程序(裂解功能噬菌体估算器)以促进对噬菌体的DLS研究。我们得出结论,DLS为学术和商业环境中噬菌体制剂的质量控制提供了一种快速、便捷且无损的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4f13/10726995/333d08efc4a2/pgad406f1.jpg

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