title: Conjugation of lentivirus to paramagnetic particles via nonviral proteins allows efficient concentration and infection of primary acute myeloid leukemia cells.
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.
authors.affiliation: King's College London, Department of Haematological and Molecular Medicine, The Rayne Institute, UK.
authors.forename: Lucas
authors.initials: L
authors.lastname: Chan
authors.forename: Darren
authors.initials: D
authors.lastname: Nesbeth
authors.forename: Taylor
authors.initials: T
authors.lastname: Mackey
authors.forename: Joanna
authors.initials: J
authors.lastname: Galea-Lauri
authors.forename: Joop
authors.initials: J
authors.lastname: Gäken
authors.forename: Francisco
authors.initials: F
authors.lastname: Martin
authors.forename: Mary
authors.initials: M
authors.lastname: Collins
authors.forename: Ghulam
authors.initials: G
authors.lastname: Mufti
authors.forename: Farzin
authors.initials: F
authors.lastname: Farzaneh
authors.forename: David
authors.initials: D
authors.lastname: Darling
mesh_terms: 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
chemical_list: D008565:Membrane Proteins; D019204:GTP-Binding Proteins