Facile lipid nanoparticle size engineering approach via controllable fusion induced by depletion forces

Lipid nanoparticles (LNPs) are among the most promising drug delivery carriers in research and development, with one major clinical application being messenger RNA (mRNA) vaccine. Current LNP production methods have the limit of generating low polydispersity index (PDI; PDI < 0.1) only for relatively small particles (<100 nm). It is known that larger LNPs have desirable properties, for example, particles with diameters in the 100 to 200 nm range have good immunogenicity. Yet, these larger particles' large PDI limits their clinical translation because of concerns about manufacturing reproducibility and possible side effects. We report here a facile approach to produce large and monodisperse (100-200 nm, PDI < 0.1) LNPs. The approach is based on adding 10 kDa polyethylene glycol (PEG) to a solution containing smaller LNPs. We show that PEG-induced depletion forces lead to the fusion of LNPs to form particles of approximately double the original size while keeping the same starting PDI. We discuss the fusion mechanism and show the parameters it depends on. In particular, we show that the fusion leads to a decrease in the fraction of empty LNPs. We show that the purification for PEG after fusion is simple and complete, thus, we believe that this is a method for the production of large LNP with low PDI that has a lot of potential to find industrial use.