Streptavidin paramagnetic particles provide a choice of three affinity-based capture and magnetic concentration strategies for retroviral vectors

Mol Ther. 2001 Apr;3(4):623-30. doi: 10.1006/mthe.2001.0268.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • Cross-Linking Reagents
  • DNA-Binding Proteins
  • Fibronectins
  • Lectins
  • PG13 protein, Nicotiana tabacum
  • Plant Proteins
  • Protein Synthesis Inhibitors
  • Succinimides
  • Puromycin
  • Biotin
  • Streptavidin