An Accessible and Scalable Pipette-Driven Microfluidic Platform for Lipid Nanoparticle Formulation

Abstract

Advances in lipid nanoparticle (LNP) technology have increased the need for accessible and scalable methods that accelerate low-volume formulation screening. Here, we describe a disposable microfluidic mixer actuated by a positive-displacement pipette equipped with dual-chamber pipette tip, providing volumetric control at a fixed flow rate ratio without external microfluidic pumps. Computational fluid dynamics analysis predicted shorter micromixing times in the high-shear spiral channel than in the low-shear design. Across three benchmark lipid formulations, the high-shear mixer reproducibly generated smaller LNPs with higher mRNA encapsulation efficiency than the low-shear mixer or manual pipetting. Orthogonal characterization further revealed higher particle concentrations, structural features consistent with compact particles, more uniform mRNA loading, and a lower empty-particle fraction. LNPs prepared using the selected high-shear mixer mediated functional mRNA delivery in 2D cell cultures, a 3D human liver tissue model, and mice. Integration with a liquid-handling robot enabled automated screening of formulation parameters. Dimensionless mixing-time analysis guided enlargement of the mixer geometry for a separate production system, which produced 75 mL preparations at 150 mL/min with particle size and mRNA encapsulation efficiency comparable to those obtained with the small-scale mixer. This platform provides an accessible, automation-compatible approach to LNP formulation screening and a route to larger-scale production.