Abstract

AbstractLipid nanoparticles have become a promising agent for biocompatible, effective, stable, and clinically approved delivery system that protect the nucleic acid from degradation while allowing cellular uptake and release of nucleic acid. Herein, we report an advanced microfluidic device and system for synthesis of uniform shape and size of mRNA encapsulated LNPs. The device consists of five repeated donut ring structures with two inlets and one outlet microfluidic channels which are assigned for organic phase solution, aqueous phase solution, and LNP recovery, respectively. It also has a minimum microfluidic channel width of 100 μm and can produce LNPs with approximately diameter from 70 to 190 nm. These LNPs can be produced less than 2 min after loading of reagents in the instrument. To verify the practical applicability of the as-developed instrument, mRNA expressing enhanced green fluorescence protein (EGFP) is encapsulated in LNPs. The gene expression of mRNA encapsulated LNPs confirmed the mechanical and chemical stability of the LNPs and its capability of gene delivery. Based on the results, the microfluidic device and system can provide a simple, fast, reliable, easy-to-use, cost-effective, and reproducible way to be applicable in LNP-based gene therapies and vaccines for treating various diseases.

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