Conjugation of lentivirus to paramagnetic particles via nonviral proteins allows efficient concentration and infection of primary acute myeloid leukemia cells
| Autor: | Lucas Chan, Taylor MacKey, Mary Collins, Farzin Farzaneh, Francisco Martin, Joanna Galea-Lauri, Darren N. Nesbeth, Ghulam J. Mufti, David Darling, Joop Gaken |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
Genetic Vectors
Immunology Virus Replication Microbiology Vesicular stomatitis Indiana virus Cell Line Viral vector Magnetics Gene Delivery Transduction (genetics) GTP-Binding Proteins Virology Murine leukemia virus Tumor Cells Cultured medicine Humans biology Lentivirus Gene Transfer Techniques Membrane Proteins Myeloid leukemia biology.organism_classification medicine.disease Molecular biology Microspheres Leukemia Myeloid Acute Leukemia Membrane protein Vesicular stomatitis virus Insect Science Biotinylation |
| DOI: | 10.1128/jvi.79.20.13190-13194.2005 |
| Popis: | 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 109CFU/ml for vesicular stomatitis virus G protein and 5 × 108for amphotropic murine leukemia virus envelope). This method also uses newly introduced membrane proteins (B7.1 and ΔLNGFR) 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. |
| Databáze: | OpenAIRE |
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