Direct plasmid DNA encapsulation within PLGA nanospheres by single oil-in-water emulsion method

Plasmid DNA was encapsulated within poly(d,l-lactic-co-glycolic acid) (PLGA) nanospheres by using polyethylene glycol (PEG) assisted solubilization technique of plasmid DNA in organic solvents. Plasmid DNA was solubilized in an organic solvent mixture composed of 80% methylene chloride and 20% DMSO by producing PEG/DNA nano-complexes having an average diameter less than 100 nm. DNA could be solubilized in the organic solvent mixture to a greater extent with increasing the weight ratio of PEG/DNA. PLGA nanospheres encapsulating DNA were successfully prepared by the single O/W emulsion method. They exhibited greater loading efficiency and better structural integrity, compared to those prepared by the W/O/W double emulsion method. Plasmid DNA could be successfully delivered to macrophage cells to express an exogenous gene. This new formulation enabled high loading of intact plasmid DNA within PLGA nanospheres useful for DNA vaccines.