通过全面优化改进 BaEV 慢病毒的生产。

IF 2.2 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Journal of virological methods Pub Date : 2024-12-28 DOI:10.1016/j.jviromet.2024.115106
Jinxue Li, Wenqiang Xu, Shengtao Luo, Hairong Zhang, Xueliang Qiu, Hao Zhang, Zhichao Liu, Qingxiao Pang
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引用次数: 0

摘要

随着细胞和基因治疗行业的快速发展,对能够有效感染不同目的细胞的慢病毒载体的需求日益增加。BaEV慢病毒已被证明能有效感染造血干细胞、原代B细胞和NK细胞,而传统的VSV-G慢病毒无法感染这些细胞。然而,在BaEV慢病毒的生产中存在着病毒产量低的问题。质粒转染后发生合胞体的形成和细胞凋亡,导致病毒产量减少。本研究全面解决了BaEV慢病毒产量低的问题。通过增加接种细胞密度、减少BaEV质粒用量、调整收获时间,BaEV慢病毒的最大滴度达到4.43E+06 IU/ml,比未优化条件下提高了369倍。进一步优化慢病毒液的纯化方法,慢病毒感染滴度达到1.00E+08 IU/ml,比优化前提高了10300倍。
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Improving the production of BaEV lentivirus by comprehensive optimization.

With the rapid development of the cell and gene therapy industry, there is an increasing demand for lentiviral vectors that can efficiently infect cells of different purposes. BaEV lentiviruses have been shown to efficiently infect hematopoietic stem cells, primary B cells, and NK cells, which traditional VSV-G lentiviruses cannot infect. However, there is a problem of low virus yield in the production of BaEV lentivirus. The formation of syncytium and apoptosis occur after plasmid transfection, and resulting in a reduction in virus production. In this study, the issue of low production of BaEV lentivirus was comprehensively improved. By increasing the cell density of inoculation, reducing the amount of BaEV plasmid, and adjusting the harvest time, the maximum titer of BaEV lentivirus reached 4.43E+ 06 IU/ml, representing an increase of 369 times compared to the unoptimized condition. Further, the purification method of lentivirus solution was optimized, and the infection titer of lentivirus reached 1.00E+ 08 IU/ml, which is 10300 times higher than pre-optimization levels.

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来源期刊
CiteScore
5.80
自引率
0.00%
发文量
209
审稿时长
41 days
期刊介绍: The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.
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