Single chain Fv: a ligand in receptor-mediated gene delivery

Gene Therapy
S GuptaP B Davis

Abstract

We have used an anti-human polymeric immunoglobulin receptor (pIgR) single chain Fv (scFv) to deliver reporter genes to epithelial cells in vitro. The scFv was constructed from a monoclonal antibody directed against pIgR and a cysteine residue was added at the carboxyl end to facilitate its conjugation to polylysine (polyK) via the heterobifunctional cross-linker SPDP. ScFv-cys was expressed in Drosophila S2 cells and purified to homogeneity using conventional column chromatography. ScFv-polyK, and polyK as control, were condensed with a DNA expression plasmid containing the luciferase reporter gene driven by the CMV promoter into unimolecular (with respect to DNA) complexes under high salt conditions. Target cells were MDCK cells transfected with human pIgR and repeatedly sorted for high-level receptor expression, with untransfected MDCK cells as control. Receptor-bearing MDCK cells were readily transfected by scFv-cys containing, pIgR directed complexes, and expression could be blocked by addition of excess human secretory component (SC), the extracellular portion of pIgR. In contrast, MDCK cells that did not express pIgR were not transfected. Nontargeted complexes were not effective in transfecting MDCK cells with or without...Continue Reading

References

Oct 21, 1988·Science·R E BirdM Whitlow
Feb 1, 1995·The Journal of Clinical Investigation·T FerkolP B Davis
Apr 26, 1994·Proceedings of the National Academy of Sciences of the United States of America·J C PeralesR W Hanson
Nov 1, 1993·The Journal of Clinical Investigation·T FerkolP B Davis
Nov 1, 1996·American Journal of Respiratory and Critical Care Medicine·P B DavisM W Konstan
Jul 29, 1999·American Journal of Respiratory Cell and Molecular Biology·E A EckmanT W Ferkol

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Citations

Aug 23, 2003·American Journal of Respiratory Cell and Molecular Biology·Sanhita GuptaPamela B Davis
Jul 5, 2005·American Journal of Respiratory Cell and Molecular Biology·Sanhita GuptaPamela B Davis
May 5, 2007·Expert Opinion on Biological Therapy·Stefanos TheoharisPeng H Tan
Feb 23, 2010·Journal of Controlled Release : Official Journal of the Controlled Release Society·Qingmin ChenDinah W Y Sah

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