病毒学
辅助病毒
重组DNA
生物
细胞培养
腺相关病毒
病毒
效价
遗传增强
病毒载体
分子生物学
病毒复制
载体(分子生物学)
基因
遗传学
作者
Zion Lee,Min Lü,Eesha Irfanullah,Morgan Soukup,Daniel Schmidt,Wei‐Shou Hu
出处
期刊:Human Gene Therapy
[Mary Ann Liebert]
日期:2023-02-01
卷期号:34 (3-4): 162-170
被引量:2
摘要
An important quality attribute of a recombinant adeno-associated virus (rAAV) as a therapeutic vector is its infectivity. Current assays to quantify infectious rAAV rely on coinfection with a helper virus such as adenovirus (Ad), which requires helper virus preparation and introduces additional variability. A simple method that has high sensitivity and removes the need for helper virus would improve assay consistency and facilitate high-throughput applications such as rAAV producer cell line development. In this study, we describe a stable assay cell line that was generated by integrating the coding sequences for AAV Rep68 and Ad E4orf6 and DNA binding protein under the control of inducible promoters. The Rep68 protein expression was further modulated by a ligand-responsive destabilization domain. In several benchmarks, the cell line gave comparable titers with those obtained using a classical Ad coinfection method. The cell line was also used to titer vectors of multiple rAAV serotypes. This cell line has the potential to serve as an effective and robust tool for quantifying infectious rAAV titers to advance gene therapy vector biomanufacturing.
科研通智能强力驱动
Strongly Powered by AbleSci AI