Abstract
Recombinant retroviruses are among several virus vectors currently being tested in clinical trials for purposes of gene therapy. As the number of clinical studies increases, accurate quantitation of retrovirus stocks and comparisons between different laboratories and clinical trials will become increasingly important. However, work from our laboratory as well as others has shown that the most commonly used quantitative retrovirus measures (i.e., titer and multiplicity of infection [MOI]) cannot be used to make accurate comparisons (1–3).
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References
Kahn M. L., Lee S. W., and Dichek D. A. (1992) Optimization of retroviral vector-mediated gene transfer into endothelial cells in vitro. Circ. Res. 71, 1508–1517.
Morgan J. R., LeDoux J. M., Snow R. G., Tompkins R. G., and Yarmush M. L. (1995) Retrovirus infection: effect of time and target cell number. J. Virol. 69, 6994–7000.
Haas D. L., Case S. S., Crooks G. M., and Kohn D. B. (2000) Critical factors influencing stable transduction of human CD34(+) cells with HIV-1-derived lentiviral vectors. Mol. Ther. 2, 71–80.
Kotani H., Newton P.B.I., Zhang S., et al. (1994) Improved methods of retroviral vector transduction and production for gene therapy. Hum. Gene Ther. 5, 19–28.
Paul R. W., Morris D., Hess B. W., Dunn J., and Overell R. W. (1993) Increased viral titer through concentration of viral harvests from retroviral packaging cell lines. Hum. Gene Ther. 4, 609–615.
Chuck A. S. and Palsson B. O. (1996) Membrane adsorption characteristics determine the kinetics of flow-through transductions. Biotech. Bioeng. 51, 260–270.
LeDoux J. M., Davis H. E., Yarmush M. L., and Morgan J. R. (1999) Kinetics of retrovirus production and decay. Biotech. Bioeng. 63, 654–662.
Andreadis S., Lavery T., Davis H. E., et al. (2000) Toward a more accurate quantitation of the activity of recombinant retroviruses: alternatives to titer and multiplicity of infection. J. Virol. 74, 3431–3439.
Shoup D. and Szabo A. (1982) Chronoamperometric current at finite disk electrodes. J. Electroanal. Chem. 140, 237–245.
Kahn M. L., Lee S. W., and Dichek D. A. (1992) Optimization of retroviral vector-mediated gene transfer into endothelial cells in vitro. Circ. Res. 71, 1508–1517.
Chuck A. C., Clarke M. F., and Palsson B. O. (1996) Retroviral infection is limited by brownian motion. Hum. Gene Ther. 7, 1527–1534.
Miller D. G., Adam M. A., and Miller A. D. (1990) Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection. Mol. Cell. Biol. 10, 4239–4242.
Salmeen I., Rimai L., Luftig R. B., et al. (1976) Hydrodynamic diameters of murine mammary, rous sarcoma and feline leukemia RNA tumor viruses: studies by laser beat frequency light-scattering spectroscopy and electron microscopy. J. Virol. 17, 584–596.
Andreadis S., Brott D. A., Fuller A. O., and Palsson B. O. (1997) Moloney murine leukemia virus-derived retroviral vectors decay intracellularly with a halflife in the range of 5.5 to 7.5 hours. J. Virol. 71, 7541–7548.
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© 2002 Humana Press Inc.
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Andreadis, S.T., Morgan, J.R. (2002). Quantitative Measurement of the Concentration of Active Recombinant Retrovirus. In: Morgan, J.R. (eds) Gene Therapy Protocols. Methods in Molecular Medicine, vol 69. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-141-8:161
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DOI: https://doi.org/10.1385/1-59259-141-8:161
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-723-6
Online ISBN: 978-1-59259-141-1
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