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Development of microfluidic process for DNA incorporation into cationic liposomes for gene therapy and vaccination

Abstract

This project focuses on the technological development of a microfluidc process for non-viral vector production, based on the electrostatic complexation between DNA and cationic liposomes for gene therapy and vaccination. The microfluidic process will be compared to the conventional bulk process. In this last process, stirred tank reactors or vortex systems can be used, but the difficult for particle size control reflects the variations in biological results and colloidal stability. The microfluidic process uses devices that process or manipulate small (10-9 to 10-18 liters) amounts of fluids, allowing the eletroctrostatic complexation in steady state flux, controlling the diffusion conditions. The liposomes will be composed by egg phosphatidylcholine, L-alpha-dioleoyl phosphatidylethanolamine and 1,2-dioleoyloxy-3-trimethylammonium propane and the pGL3-Control DNA will be used as control. The cationic liposome/DNA complexes obtained by microfluidic and bulk processes will be characterized according their physico-chemical properties zeta potential, morphology, mean hydrodynamic diameter, polydispersity and size distribution. Transfections in vitro will be the biological evaluation. Our previous experience with cationic liposomes focused the development of a DNA vaccine against tuberculosis (one patent, one addition certificate and 4 publications) and scale up studies for liposome production and its complexation with DNA based on bulk processe (FAPESP PIPE II). Furthermore, the "empty" cationic liposome production using microfluidic device is also in development and it is afforded by Jovem Pesquisador project (Nanotecnologia/CNPq), with a master's student at the end of her research. In this context, this project will contribute to the development of a new feasible process, allowing the development of new pharmaceutical products for gene and vaccine therapies. (AU)

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VEICULO: TITULO (DATA)
VEICULO: TITULO (DATA)