Characterization of diverse viral vector preparations, using a simple and rapid whole-virion dot-blot method.
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 vectors, 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.
Nelson
D M
DM
Clinical Gene Therapy Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892-1851, USA.
Wahlfors
J J
JJ
Chen
L
L
Onodera
M
M
Morgan
R A
RA
D000256:Adenoviridae / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D017403:In Situ Hybridization / Q000379:methods*; D013539:Simian virus 40 / Q000235:genetics; D012845:Sindbis Virus / Q000235:genetics; D014773:Virology / Q000379:methods*