Conjugation of lentivirus to paramagnetic particles via nonviral proteins allows efficient concentration and infection of primary acute myeloid leukemia cells

被引:32
作者
Chan, L
Nesbeth, D
MacKey, T
Galea-Lauri, J
Gäken, J
Martin, F
Collins, M
Mufti, G
Farzaneh, F
Darling, D
机构
[1] Kings Coll London, Dept Haematol & Mol Med, Rayne Inst, London SE5 9NU, England
[2] UCL, Windeyer Inst Med Sci, Dept Immunol & Mol Pathol, London, England
关键词
D O I
10.1128/JVI.79.20.13190-13194.2005
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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 108 for amphotropic murine leukemia virus envelope). This method also uses newly introduced membrane proteins (B7.1 and Delta LNGFR) 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.
引用
收藏
页码:13190 / 13194
页数:5
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