The First Micromixing Method Delivering Consistent Lipid Nanoparticle Formulations Seamlessly Across All Scales

The development of RNA-based vaccines and therapies using lipid nanoparticles (LNPs) is hindered by the absence of a unified formulation technology that covers the entire development pipeline – from microliter-scale screening to tens-of-liters manufacturing. This gap requires transitions between different formulation techniques during scale-up, such as microfluidics, impingement jet mixing (IJM) or T-mixing, which can affect key LNP quality parameters, including size, polydispersity index (PDI), encapsulation efficiency (EE%), and morphology. Current approaches lead to prolonged, expensive, and sometimes unsuccessful LNP-based drug development. Here, we introduce NanoPULSE, a novel, patented active micromixing technology that ensures consistent LNP characteristics across all production volumes. Lagrangian numerical simulations are used to predict the impact of input parameters on particle characteristics and optimize the mixing channel’s dimensions. Simulation results permitted the introduction of a novel index to quantify the mixing efficiency of NanoPULSE. Multi-angle dynamic light scattering (MADLS) measurements confirm that empty and RNA-loaded LNPs maintain constant sizes over production volumes ranging from 500 µL to 4 L, with PDIs remaining below 0.2, validating the seamless scalability of the technology. The EE%, yield and structural morphology of RNA-LNPs produced with NanoPULSE are characterized at different scales and benchmarked with standard microfluidic approaches. By modulating the solvents’ injection periods, NanoPULSE can produce LNPs of variable sizes using the same formulation. The LNPs sizes and PDIs measured at different input parameters are correlated with simulations, linking the mixing performance to the nanoparticle characteristics. These findings underscore the promising repeatability, scalability, and efficiency of LNP synthesis via NanoPULSE.