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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

Mol ther

Publication Date

04/2001

Volume

3

Pages

623 - 630

Keywords

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