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अमूर्त

Characterization of Human Serum Albumin-Facilitated Lipofection Gene Delivery Strategy

Robert W Arpke and Pi-Wan Cheng

We report the characterization of a facilitated lipofection strategy, which employs a formulation prepared with human serum albumin, DMRIE-C and pCMV?. The transfection complexes in the lipofection formulation containing albumin were characterized for size by light scattering, intracellular trafficking by confocal microscopy, and uptake mechanism by inhibitors. Supplementation of the formulation of DMRIE-C plus pCMV? with albumin enhances lipofection efficiency 8-9 fold and the size of the complexes 2-2.5 fold as measured by light scattering. Analysis of intracellular trafficking of the transfection complexes by confocal microscopy reveals colocalization of DNA and albumin in the cytoplasm and the nucleus. Pretreatment of the cells with chlorpromazine or excess albumin, cytochalasin B or filipin complexes results in inhibition of the transfection efficiency by 20%, 55%, or none, respectively. The results suggest a moderate involvement of clathrin, a significant involvement of actinassociated macropinocytosis, and no involvement of caveolae in the uptake of the transfection complexes prepared with human serum albumin, DMRIE-C and pCMV?.