Continuous Production of Discrete Plasmid DNA-Polycation Nanoparticles Using Flash Nanocomplexation

Area of Science:

• Biomaterials Science
• Nanotechnology
• Gene Therapy

Background:

• Linear polyethyleneimine (lPEI) is a proven gene carrier for DNA, RNA, and mRNA.
• Current methods for producing lPEI/nucleic acid nanoparticles face scalability issues, hindering clinical applications.
• lPEI/DNA nanoparticles are crucial for advancing gene medicine but require scalable manufacturing.

Purpose of the Study:

• To develop a scalable and continuous method for producing lPEI/plasmid DNA nanoparticles.
• To address the limitations of traditional batch methods in nanoparticle manufacturing.
• To enable efficient clinical translation of lPEI/DNA nanoparticle-based gene therapies.

Main Methods:

• A flash nanocomplexation (FNC) method utilizing a confined impinging jet device.
• Rapid, dynamic, and homogeneous mixing for complex coacervation of DNA and lPEI.
• Continuous production of polyelectrolyte complex nanoparticles.

Main Results:

• FNC achieved continuous production of lPEI/plasmid DNA nanoparticles with narrow size and shape distribution.
• The number of DNA plasmids packaged per nanoparticle was comparable to small-scale batch methods.
• Nanoparticles produced via FNC demonstrated similar cell transfection efficiency to batch-produced ones.

Conclusions:

• Flash nanocomplexation (FNC) is an effective and scalable method for producing well-controlled lPEI/plasmid DNA nanoparticles.
• The FNC method overcomes scalability challenges, facilitating the clinical translation of gene therapies.
• This continuous manufacturing approach ensures consistent quality for nanoparticle-based gene medicine.