Conjugation of lentivirus to paramagnetic particles via nonviral proteins allows efficient concentration and infection of primary acute myeloid leukemia cells.
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.
Chan
Lucas
L
King's College London, Department of Haematological and Molecular Medicine, The Rayne Institute, UK.
Nesbeth
Darren
D
Mackey
Taylor
T
Galea-Lauri
Joanna
J
Gäken
Joop
J
Martin
Francisco
F
Collins
Mary
M
Mufti
Ghulam
G
Farzaneh
Farzin
F
Darling
David
D
D002460:Cell Line; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000502:physiology*; D015470:Leukemia, Myeloid, Acute; D008280:Magnetics; D008565:Membrane Proteins / Q000378:metabolism*; D008863:Microspheres; D014407:Tumor Cells, Cultured / Q000821:virology; D014721:Vesicular stomatitis Indiana virus / Q000737:chemistry; D014779:Virus Replication
D008565:Membrane Proteins; D019204:GTP-Binding Proteins