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Three strategies have been designed to concentrate infectious retroviral vectors from the supernatants of human- (HT1080) and murine- (NIH 3T3) based packaging cells. Streptavidin-conjugated paramagnetic particles in conjunction with (i) antibodies directed against murine fibronectin, (ii) biotinylated lectins, or (iii) biotin-modified packaging cell-surface proteins allow affinity-mediated magnetic concentration of retroviral vectors. Retroviral titers (assayed by colony formation of human myeloid K562 cells) are increased by 1-4 x 10(3)-fold after volume reductions of only 125-fold. Using these procedures, preparations of 5 x 10(8) cfu/ml are routinely made from relatively low-titer (2-5 x 10(5) cfu/ml) starting material. High-titer (paramagnetic) retroviral vector preparations can be used for magnetic field-dependent retroviral infection in vitro. Magnetic field-dependent localization such as this may enable the in vivo administration of formulations that concentrate retroviral infection to the required target tissues and organs.

Original publication

DOI

10.1006/mthe.2001.0268

Type

Journal article

Journal

Molecular Therapy

Publication Date

04/2001

Volume

3

Pages

623 - 630

Addresses

Guy's, King's, and St. Thomas' School of Medicine, Department of Molecular Medicine, The Rayne Institute, 123 Coldharbour Lane, London, SE5 9NU, United Kingdom.

Keywords

Cell Line, Hela Cells, K562 Cells, 3T3 Cells, Animals, Humans, Mice, Retroviridae, Puromycin, Succinimides, Biotin, Streptavidin, DNA-Binding Proteins, Fibronectins, Lectins, Plant Proteins, Protein Synthesis Inhibitors, Cross-Linking Reagents, Microscopy, Fluorescence, Gene Transfer Techniques, Genetic Vectors, Magnetics