Streptavidin paramagnetic particles provide a choice of three affinity-based capture and magnetic concentration strategies for retroviral vectors.
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.
Hughes
C
C
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.
Galea-Lauri
J
J
Farzaneh
F
F
Darling
D
D
D016475:3T3 Cells; D000818:Animals; D001710:Biotin / Q000378:metabolism; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000235:genetics; D005353:Fibronectins / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D037102:Lectins / Q000235:genetics; D008280:Magnetics; D051379:Mice; D008856:Microscopy, Fluorescence; D010940:Plant Proteins / Q000235:genetics; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011691:Puromycin / Q000494:pharmacology; D012190:Retroviridae / Q000235:genetics*; D019809:Streptavidin / Q000737:chemistry*; D013388:Succinimides / Q000494:pharmacology
D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D005353:Fibronectins; D037102:Lectins; C089914:PG13 protein, Nicotiana tabacum; D010940:Plant Proteins; D011500:Protein Synthesis Inhibitors; D013388:Succinimides; D011691:Puromycin; D001710:Biotin; D019809:Streptavidin