Optimized production and concentration of lentiviral vectors containing large inserts
Optimized production and concentration of lentiviral vectors containing large inserts
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DOI：
10.1002/jgm.1052
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发表时间：
2007-07-01
影响因子：
3.5
通讯作者：
Foerster, John
中科院分区：
文献类型：
--
作者：
Al Yacoub, Nadya;Romanowska, Malgorzata;Foerster, John
Generation of high titer lentiviral stocks and efficient virus concentration are central to maximize the utility of lentiviral technology. Here we evaluate published protocols for lentivirus production on a range of transfer vectors differing in size (7.5-13.2 kb). We present a modified virus production protocol robustly yielding useful titers (Up to 10(7)/ml) for a range of different transfer vectors containing packaging inserts up to 7.5 kb. Moreover, we find that virus recovery after concentration by ultracentrifugation depends on the size of the packaged inserts, heavily decreasing for large packaged inserts. We describe a fast (4 h) centrifugation protocol at reduced speed allowing high virus recovery even for large and fragile lentivirus vectors. The protocols outlined in the current report should be useful for many labs interested in producing and concentrating high titer lentiviral stocks. Copyright (C) 2007 John Wiley & Sons, Ltd.
