This study investigates coaxial electrohydrodynamic spraying (electrospray for short) as a novel, rapid, real time and single-step method to produce oligodeoxynucleotide (ODN) encapsulated lipoplex nanoparticles for either intravenous injection or, potentially, pulmonary delivery. Using a coaxial needle setup, we produced G3139 (oblimerson sodium, or Genasense) encapsulated lipoplex nanoparticles, and investigated the effects of production parameters on nanoparticle size and structure. Careful control of production parameters yielded lipoplex nanoparticles 190 +/- 39 nm in diameter with unilamellar structure and 90 +/- 6% encapsulation efficiency of G3139. Both nontargeted and transferrin-targeted G3139 lipoplex nanoparticles were efficiently delivered to K562 cells and downregulated the bcl-2 protein expression by 34 +/- 6% and 57 +/- 3% respectively.