Increased viral titer through concentration of viral harvests from retroviral packaging lines.
Dependent on the viral vector and the specific assay used, viral titers produced from commonly used retroviral packaging cell lines have an upper limit in the range of 10(5) to 10(7) infectious units/ml. We have developed a generally applicable method, using hollow-fiber filtration technology, which allows for the concentration of infectious virus derived from packaging lines. This method resulted in a reproducible 10- to 30-fold increase in viral titer and can readily be scaled to accommodate larger input volumes. Over 80% of the input virus is recovered in an infectious form in the concentrate. Concentrated virus containing media was seen to produce higher infection frequencies in Jurkat T cells as compared to unconcentrated virus containing media; however, this was not proportional to the differences in viral titer observed by limiting dilution analysis on NIH-3T3 cells. These results are discussed in relation to the importance of factors other than viral titer in determining transduction frequencies.
Paul
R W
RW
Targeted Genetics Corporation, Seattle, WA 98101.
Morris
D
D
Hess
B W
BW
Dunn
J
J
Overell
R W
RW
D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D005821:Genetic Techniques; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D012190:Retroviridae / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D013601:T-Lymphocytes / Q000382:microbiology; D014776:Virus Cultivation
D012194:RNA-Directed DNA Polymerase