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Nonviral producer cell proteins incorporated into retroviral vector surfaces profoundly influence infectivity and in vivo half-life. We report the purification and concentration of lentiviral vectors using these surface proteins as an efficient gene transduction strategy. Biotinylation of these proteins and streptavidin paramagnetic particle concentration enhances titer 400- to 2,500-fold (to 10(9) CFU/ml for vesicular stomatitis virus G protein and 5 x 10(8) for amphotropic murine leukemia virus envelope). This method also uses newly introduced membrane proteins (B7.1 and DeltaLNGFR) directed to lentiviral surfaces, allowing up to 17,000-fold concentrations. Particle conjugation of lentivirus allows facile manipulation in vitro, resulting in the transduction of 48 to 94% of human acute myeloid leukemia blasts.

Original publication

DOI

10.1128/jvi.79.20.13190-13194.2005

Type

Journal article

Journal

Journal of virology

Publication Date

10/2005

Volume

79

Pages

13190 - 13194

Addresses

King's College London, Department of Haematological and Molecular Medicine, The Rayne Institute, UK.

Keywords

Cell Line, Tumor Cells, Cultured, Humans, Lentivirus, GTP-Binding Proteins, Membrane Proteins, Gene Transfer Techniques, Microspheres, Virus Replication, Genetic Vectors, Magnetics, Vesicular stomatitis Indiana virus, Leukemia, Myeloid, Acute