The contribution of poly-L-lysine, epidermal growth factor and streptavidin to EGF/PLL/DNA polyplex formation.

High-level targeted gene delivery has been demonstrated by molecular conjugates in vitro; however, in vivo delivery has been limited. The complexity of the resulting protein/DNA polyplex and a lack of understanding of its formation are persistent limitations. In this report, we show the effect of the DNA-binding agent poly-L-lysine (PLL), the ligand epidermal growth factor (EGF), and the coupling protein streptavidin on particle size, charge and gene delivery. Smaller (< 80 nm) and more stable polyplexes were obtained with PLL1116 than with shorter versions of PLL, especially in 0.15 M NaCl. Stability was increased by adding streptavidin to the polyplex; however, EGF increased particle size (> 1000 nm) and decreased gene delivery when > 300 EGF molecules per polyplex were used, indicating that a critical number of EGF molecules was needed for efficient gene delivery. The correct combination of these components resulted in the most efficient gene delivery in vitro and now provide for testing a more stable protein/DNA polyplex to aid in enhancing gene delivery in vivo.