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Conjugation of lentivirus to paramagnetic particles via nonviral proteins allows efficient concentration and infection of primary acute myeloid leukemia cells.
- Source :
-
Journal of virology [J Virol] 2005 Oct; Vol. 79 (20), pp. 13190-4. - Publication Year :
- 2005
-
Abstract
- 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.
- Subjects :
- Cell Line
GTP-Binding Proteins genetics
GTP-Binding Proteins metabolism
Genetic Vectors genetics
Humans
Lentivirus genetics
Tumor Cells, Cultured virology
Vesicular stomatitis Indiana virus chemistry
Virus Replication
Gene Transfer Techniques
Genetic Vectors physiology
Lentivirus physiology
Leukemia, Myeloid, Acute
Magnetics
Membrane Proteins metabolism
Microspheres
Subjects
Details
- Language :
- English
- ISSN :
- 0022-538X
- Volume :
- 79
- Issue :
- 20
- Database :
- MEDLINE
- Journal :
- Journal of virology
- Publication Type :
- Academic Journal
- Accession number :
- 16189021
- Full Text :
- https://doi.org/10.1128/JVI.79.20.13190-13194.2005