Improving the production of BaEV lentivirus by comprehensive optimization

• Published in: Journal of virological methods 2025-04, Vol.333, p.115106, Article 115106
• Main Authors: Li, Jinxue, Xu, Wenqiang, Luo, Shengtao, Zhang, Hairong, Qiu, Xueliang, Zhang, Hao, Liu, Zhichao, Pang, Qingxiao
• Format: Article
• Language: English
• Subjects: BaEV-LV; CAR-NK; Titer; Transfection; Ultracentrifugation
• ISSN: 0166-0934; 1879-0984; 1879-0984
• DOI: 10.1016/j.jviromet.2024.115106

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. •BaEV lentivirus packing titers raised to 4.43E+ 06 IU/ml, which increased 369 times compared to the unoptimized condition.•BaEV purification titers reached 1.00E+ 08 IU/ml, which is 10300 times higher than pre-optimization levels.•The formation of syncytium and the lysis of packing cells were effectively alleviated.