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Scalable Purification of Plasmid DNA Nanoparticles by Tangential Flow Filtration for Systemic Delivery
Heng-Wen Liu1,2, Yizong Hu2,3, Yong Ren1,2
1Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, Maryland 21218, United States.
ACS Applied Materials & Interfaces
|June 24, 2021
Summary
We developed a scalable purification method for plasmid DNA (pDNA) nanoparticles using tangential flow filtration (TFF). This process reduces toxicity by removing excess linear polyethylenimine (lPEI) while maintaining gene delivery efficiency for clinical applications.
Area of Science:
- Biotechnology
- Nanomedicine
- Gene Therapy
Background:
- Plasmid DNA (pDNA) complexed with linear polyethylenimine (lPEI) are effective non-viral gene delivery vectors.
- Scalability, reproducibility, and toxicity are key challenges for clinical translation of pDNA/lPEI nanoparticles.
Purpose of the Study:
- To develop a scalable purification method for pDNA/lPEI nanoparticles.
- To improve the biocompatibility of pDNA/lPEI nanoparticles by reducing uncomplexed lPEI.
- To validate the purified nanoparticles for clinical gene nanomedicine applications.
Main Methods:
- Flash Nanocomplexation (FNC) for scalable nanoparticle formulation.
- Tangential Flow Filtration (TFF) for purification and reduction of uncomplexed lPEI.
- In vitro and in vivo studies to assess toxicity and transfection efficiency.
Main Results:
- Achieved a 60% reduction in uncomplexed lPEI concentration.
- Preserved nanoparticle size and morphology after purification.
- Demonstrated significantly reduced toxicity with maintained transfection efficiency in vitro and in vivo.
- TFF enabled solvent exchange to isotonic solutions and nanoparticle concentration.
Conclusions:
- TFF is a scalable and effective purification method for pDNA/lPEI nanoparticles.
- Purified nanoparticles exhibit improved biocompatibility and reduced toxicity.
- Combining FNC and TFF offers a viable strategy for clinical translation of gene nanomedicine.
