Scalable Purification of Plasmid DNA Nanoparticles by Tangential Flow Filtration for Systemic Delivery

• DOI: 10.1021/acsami.1c05750
• Publication Date: 2021-06-24T05:31:02Z
• Authors: Heng-Wen Liu; Yizong Hu; Yong Ren; Hwanhee Nam; Jose Luis Santos; Shirley Ng; Like Gong; Mary Brummet; Christine A. Carr...; Christopher G. Ul...; Martin G. Pomper; Il Minn; Hai-Quan Mao
• Keywords: Polyethylenimine; Biocompatibility; Filtration (mathematics)

Plasmid DNA (pDNA) nanoparticles synthesized by complexation with linear polyethylenimine (lPEI) are one of the most effective non-viral gene delivery vehicles. However, lack scalable and reproducible production methods high toxicity have hindered their clinical translation. Previously, we developed a flash nanocomplexation (FNC) technique to formulate pDNA/lPEI using continuous flow process. Here, report tangential filtration (TFF)-based purification method reduce uncomplexed lPEI concentration in nanoparticle formulation improve its biocompatibility. The optimized procedures achieved 60% reduction preservation size morphology. Both vitro vivo studies showed that purified significantly reduced while maintaining transfection efficiency. TFF also allows for gradual exchange solvents isotonic solutions further concentrating injection. Combining FNC purification, validated future translation this nanomedicine.