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

TLDR

Lipid nanoparticles for RNA therapeutics must be reliably produced across development scales, yet microfluidic methods, though precise, have been limited by throughput scaling challenges. The authors introduce a scalable, parallelized microfluidic device that uses 128 simultaneous mixing channels to overcome these throughput limitations. Its mechanism relies on an array of 128 mixing channels that operate concurrently to parallelize production. The PMD delivers over 100‑fold higher production rates than single‑channel microfluidics while preserving LNP physical properties and potency, and in mice achieves 4‑fold greater hepatic gene silencing with siRNA and 5‑fold higher luciferase expression, demonstrating its suitability for scalable, reproducible RNA therapeutic and vaccine formulations.

Abstract

A major challenge to advance lipid nanoparticles (LNPs) for RNA therapeutics is the development of formulations that can be produced reliably across the various scales of drug development. Microfluidics can generate LNPs with precisely defined properties, but have been limited by challenges in scaling throughput. To address this challenge, we present a scalable, parallelized microfluidic device (PMD) that incorporates an array of 128 mixing channels that operate simultaneously. The PMD achieves a >100× production rate compared to single microfluidic channels, without sacrificing desirable LNP physical properties and potency typical of microfluidic-generated LNPs. In mice, we show superior delivery of LNPs encapsulating either Factor VII siRNA or luciferase-encoding mRNA generated using a PMD compared to conventional mixing, with a 4-fold increase in hepatic gene silencing and 5-fold increase in luciferase expression, respectively. These results suggest that this PMD can generate scalable and reproducible LNP formulations needed for emerging clinical applications, including RNA therapeutics and vaccines.