A receptor-mediated gene delivery system using streptavidin and biotin-derivatized, pegylated epidermal growth factor.
An efficient receptor-mediated non-viral gene delivery formulation based on mono-pegylated recombinant human epidermal growth factor (EGF) was developed using a streptavidin-biotin system. Biotin-derivatized and mono-pegylated EGF was prepared by conjugating a biotin-PEG-NHS derivative to EGF and purified through a chromatographic method. Luciferase plasmid DNA and polyethylenimine (PEI) were complexed to form positively charged nanoparticles on which negatively charged streptavidin was first coated and then biotin-PEG-EGF conjugate was immobilized via streptavidin-biotin interaction. The EGF-PEG-biotin-streptavidin-PEI-DNA complexes were characterized in terms of their effective diameter and surface zeta (zeta)-potential value under various formulation conditions. The formulated complexes exhibited high transfection efficiency (approximately 10(8) in luciferase activity) with no inter-particle aggregation. This was attributed to enhanced cellular uptake of the resultant complexes via receptor-mediated endocytosis. Furthermore, in the presence of serum proteins, a slight decrease in transfection efficiency was observed due to the presence of PEG chains on the surface.
Lee
Haeshin
H
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Taejon 305-701, South Korea.
Kim
Tae Hyoung
TH
Park
Tae Gwan
TG
D001710:Biotin / Q000008:administration & dosage* / Q000493:pharmacokinetics; D002626:Chemistry, Pharmaceutical; D016503:Drug Delivery Systems / Q000379:methods* / Q000706:statistics & numerical data; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology; D004815:Epidermal Growth Factor / Q000008:administration & dosage* / Q000493:pharmacokinetics; D015316:Genetic Therapy / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D010957:Plasmids / Q000008:administration & dosage / Q000493:pharmacokinetics; D011094:Polyethyleneimine / Q000008:administration & dosage* / Q000493:pharmacokinetics; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000493:pharmacokinetics; D019809:Streptavidin / Q000008:administration & dosage* / Q000493:pharmacokinetics; D014407:Tumor Cells, Cultured / Q000187:drug effects
D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; D001710:Biotin; D011094:Polyethyleneimine; D019809:Streptavidin