Fars Alanazi1, Zhen F Fu, D Robert Lu. 1. Department of Pharmaceutical and Biomedical Sciences, University of Georgia, Athens, Georgia 30602, USA.
Abstract
PURPOSE: To evaluate the transfection efficiency in cell culture of rabies plasmid DNA vaccine carried by a novel artificial lipoprotein system. METHODS: Phospholipid nanoemulsions resembling the lipid core of natural lipoproteins were prepared. The artificial lipoprotein carrier system for DNA was constructed by assembling of the nanoemulsion (NE)-palmitoyl-poly-L-lysine (p-PLL)-rabies DNA complex. Agarose gel electrophoresis, zeta potential, and mobility measurement were conducted to determine the surface charge balance in various complex compositions. Transfection and transfection efficiency were examined by fluorescence microscopy and flow cytometry, respectively. RESULTS: The artificial lipoprotein system was successfully constructed and the rabies DNA vaccine was effectively transfected in glioma cell line SF-767. The amount of p-PLL incorporated into the artificial lipoprotein formulations had a significant effect on transfection efficiency. The new system also proved to be more efficient in cellular transfection of rabies DNA vaccine than the commercial lipofectamine formulation. CONCLUSIONS: Effective transfection of rabies DNA vaccine in cell culture can be achieved using the novel artificial lipoprotein carrier system, and the charge balance of the NE-p-PLL-DNA complex appears an important factor.
PURPOSE: To evaluate the transfection efficiency in cell culture of rabies plasmid DNA vaccine carried by a novel artificial lipoprotein system. METHODS:Phospholipid nanoemulsions resembling the lipid core of natural lipoproteins were prepared. The artificial lipoprotein carrier system for DNA was constructed by assembling of the nanoemulsion (NE)-palmitoyl-poly-L-lysine (p-PLL)-rabies DNA complex. Agarose gel electrophoresis, zeta potential, and mobility measurement were conducted to determine the surface charge balance in various complex compositions. Transfection and transfection efficiency were examined by fluorescence microscopy and flow cytometry, respectively. RESULTS: The artificial lipoprotein system was successfully constructed and the rabies DNA vaccine was effectively transfected in glioma cell line SF-767. The amount of p-PLL incorporated into the artificial lipoprotein formulations had a significant effect on transfection efficiency. The new system also proved to be more efficient in cellular transfection of rabies DNA vaccine than the commercial lipofectamine formulation. CONCLUSIONS: Effective transfection of rabies DNA vaccine in cell culture can be achieved using the novel artificial lipoprotein carrier system, and the charge balance of the NE-p-PLL-DNA complex appears an important factor.
Authors: David A Bull; Stephen H Bailey; Jeffrey J Rentz; James S Zebrack; Minhyung Lee; Sheldon E Litwin; Sun Wan Kim Journal: J Control Release Date: 2003-12-05 Impact factor: 9.776