An intracellular delivery vehicle for protein transduction of micro-dystrophin

J Drug Target. 2005 Feb;13(2):81-7. doi: 10.1080/10611860400029002.

Abstract

The Fv fragment of an antibody that selectively targets and penetrates skeletal muscle in vivo was produced as a fusion protein with a micro-dystrophin for use as a delivery vehicle to transport micro-dystrophin into muscle cells. Fv-micro-dystrophin was produced as a secreted protein by transient transfection of Fv-micro-dystrophin cDNA in COS-7 cells and as a non-secreted protein by permanent transfection in Pichia pastoris. Isolated Fv-micro-dystrophin was shown to be full-length by Western blot analysis. Fv-micro-dystrophin penetrated multiple cell lines in vitro, and it localized to the plasma membrane of a cell line with membrane beta-dystroglycan. In the absence of membrane beta-dystroglycan, it localized to the cytoplasm. Antibody-mediated transduction of micro-dystrophin into muscle cells is a potential therapy for dystrophin-deficient muscular dystrophies.

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Line
  • Cell Membrane / metabolism*
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Cytoplasm / metabolism
  • Dystroglycans / metabolism
  • Dystrophin / genetics
  • Dystrophin / metabolism*
  • Humans
  • Immunoglobulin Fragments*
  • Lymphokines*
  • Myoblasts / metabolism
  • Pichia / metabolism
  • Protein Sorting Signals*
  • Protein Transport
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Sialoglycoproteins*
  • Transfection

Substances

  • Dystrophin
  • Fv protein, human
  • Immunoglobulin Fragments
  • Lymphokines
  • Protein Sorting Signals
  • Recombinant Fusion Proteins
  • Sialoglycoproteins
  • Dystroglycans