Conjugation of lentivirus to paramagnetic particles via nonviral proteins allows efficient concentration and infection of primary acute myeloid leukemia cells
Conjugation of lentivirus to paramagnetic particles via nonviral proteins allows efficient concentration and infection of primary acute myeloid leukemia cells
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DOI：
10.1128/jvi.79.20.13190-13194.2005
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发表时间：
2005-10-01
影响因子：
5.4
通讯作者：
Darling, D
中科院分区：
文献类型：
--
作者：
Chan, L;Nesbeth, D;Darling, D
掺入逆转录病毒载体表面的非病毒生产细胞蛋白深刻影响感染性和体内半衰期。我们报告使用这些表面蛋白作为有效的基因转导策略来纯化和浓缩慢病毒载体。这些蛋白质和链霉亲和素顺磁颗粒浓度的生物素化可将滴度提高 400 至 2,500 倍（水泡性口炎病毒 G 蛋白为 10(9) CFU/ml，双嗜性鼠白血病病毒包膜为 5 x 108）。该方法还使用新引入的针对慢病毒表面的膜蛋白（B7.1 和 Delta LNGFR），浓度高达 17,000 倍。慢病毒的颗粒接合可以在体外轻松操作，从而转导 48% 至 94% 的人类急性髓系白血病母细胞。
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.