Characterization of diverse viral vector preparations, using a simple and rapid whole-virion dot-blot method

被引:9
|
作者
Nelson, DM [1 ]
Wahlfors, JJ [1 ]
Chen, L [1 ]
Onodera, M [1 ]
Morgan, RA [1 ]
机构
[1] Natl Human Genome Res Inst, Gene Transfer Technol Sect, Clin Gene Therapy Branch, NIH, Bethesda, MD 20892 USA
关键词
D O I
10.1089/hum.1998.9.16-2401
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
A number of different viruses have been adapted as gene transfer vectors, including retroviruses, adenoviruses, adenoassociated viruses (AAVs), herpes simplex virus, SV40 viruses, and alphaviruses (both Semliki Forest and Sindbis viruses), One of the major rate-limiting and time-consuming steps in the characterization of these vectors is the process of determining the viral vector titers, In addition, there is no "universal" method that can be used to rapidly estimate the titer and the utility of viral vector preparations. We demonstrate here that supernatant from diverse classes of viral victors, with either RNA or DNA genomes, can be rapidly evaluated by a simple virus dot-blot hybridization without prior extraction of nucleic acids. This system can provide a reliable screen for physical titer of viral vector supernatants in 1 day.
引用
收藏
页码:2401 / 2405
页数:5
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