Scalable mRNA and siRNA Lipid Nanoparticle Production Using a Parallelized Microfluidic Device.

Area of Science:

• Biotechnology
• Nanomedicine
• Drug Delivery

Background:

• Advancing RNA therapeutics requires reliable production of lipid nanoparticles (LNPs) across development scales.
• Microfluidics offers precise LNP property control but faces throughput limitations.
• Scalable manufacturing is crucial for translating LNP-based therapies to clinical use.

Purpose of the Study:

• To develop a scalable microfluidic device for high-throughput LNP production.
• To evaluate the physical properties, potency, and in vivo delivery of LNPs produced by the device.
• To demonstrate the potential of the device for clinical applications of RNA therapeutics.

Main Methods:

• A parallelized microfluidic device (PMD) with 128 simultaneous mixing channels was designed and fabricated.
• LNPs encapsulating Factor VII siRNA or luciferase mRNA were produced using the PMD and conventional mixing methods.
• LNP physical properties, potency, and in vivo gene silencing/expression were assessed in mice.

Main Results:

• The PMD achieved over 100x higher production rate than single microfluidic channels without compromising LNP properties.
• PMD-generated LNPs showed superior in vivo delivery, with a 4-fold increase in hepatic gene silencing and 5-fold increase in luciferase expression compared to conventional mixing.
• The scalability and reproducibility of LNP formulations were demonstrated.

Conclusions:

• The scalable, parallelized microfluidic device (PMD) overcomes throughput limitations of microfluidic LNP production.
• The PMD generates reproducible and potent LNP formulations suitable for clinical applications, including RNA therapeutics and vaccines.
• This technology facilitates the advancement of LNP-based therapies from development to clinical practice.