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作为一种抗菌基因疗法，无病毒的病毒样P4-EKORhE多溶素转导颗粒的高产生物生产。

High-yield bioproduction of virus-free virus-like P4-EKORhE multi-lysin transducing particles as an antimicrobial gene therapeutic.

作者信息

Ramirez-Garcia Robert, Sagona Antonia P, Barr Jeremy J, Jaramillo Alfonso

机构信息

School of Life Sciences, Faculty of Science, Engineering and Medicine, The University of Warwick, Coventry, United Kingdom.

School of Biological Sciences, Faculty of Sciences, Monash University, Melbourne, VIC, Australia.

出版信息

Front Cell Infect Microbiol. 2025 Jun 25;15:1561443. doi: 10.3389/fcimb.2025.1561443. eCollection 2025.

DOI:10.3389/fcimb.2025.1561443
PMID:40636261
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12239096/
Abstract

A description of the construction of the bioengineered P4-EKORhE and a comprehensive method for producing very high yields (up to 10 particles per millilitre) enable the use of virus-like particles to transduce genetically encoded antimicrobials through a combination of synthetic biology and optimised upstream and downstream processing. The final product, a gene-delivered antimicrobial in the form of the multi-lysin cassette, is fully functional before and after packaging within P4-EKORhE particles. The antimicrobial activity of the multi-lysin cassette, characterised by its lysis proteins, was tested in both pure bacterial cultures and a model of phage infection in co-culture with A549 immortalised human epithelial tissue cells. This work exemplifies several bioproduction methods and demonstrates how the virology of the P4 and P2 phages can be harnessed to establish a bioprocess for producing transducing particles at very high yields, avoiding contamination by the natural virus while maintaining the antimicrobial effectiveness of the final product.

摘要

对生物工程改造的P4-EKORhE构建过程的描述以及一种实现非常高产量（每毫升高达10个颗粒）的综合方法，使得通过合成生物学与优化的上游和下游加工相结合，利用病毒样颗粒转导基因编码的抗菌剂成为可能。最终产品，即多溶素盒形式的基因递送抗菌剂，在包装于P4-EKORhE颗粒之前和之后均具有完全功能。通过其裂解蛋白表征的多溶素盒的抗菌活性，在纯细菌培养物以及与永生化人上皮组织细胞A549共培养的噬菌体感染模型中进行了测试。这项工作例证了几种生物生产方法，并展示了如何利用P4和P2噬菌体的病毒学来建立一种以非常高的产量生产转导颗粒的生物工艺，在避免天然病毒污染的同时保持最终产品的抗菌效力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/0518d7681bec/fcimb-15-1561443-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/48605ade8528/fcimb-15-1561443-g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/38e2fcb0a300/fcimb-15-1561443-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/8cb37b7f14bc/fcimb-15-1561443-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/b9956b10b86f/fcimb-15-1561443-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/0518d7681bec/fcimb-15-1561443-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/48605ade8528/fcimb-15-1561443-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/1f3715ec7270/fcimb-15-1561443-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/b6f50ee2ef6a/fcimb-15-1561443-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/aa9d39b802fb/fcimb-15-1561443-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/38e2fcb0a300/fcimb-15-1561443-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/8cb37b7f14bc/fcimb-15-1561443-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/b9956b10b86f/fcimb-15-1561443-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/126a/12239096/0518d7681bec/fcimb-15-1561443-g008.jpg

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