Probing microfluidic assembly of mRNA encapsulated lipid nanoparticles

Messenger RNA medicines enables the treatment of classically undruggable diseases and immunisation against emergent pathogens by administering the genetic sequence encoding for specific proteins or antigens. However naked mRNA is highly susceptible to endogenous nucleases and does not associate with cells, hence must be formulated. The most translatable approach is to employ small (50-150 nm diameter) lipid nanoparticles (LNPs) which protect and facilitate cellular transport of mRNA. The physicochemical nanostructure of the LNPs are critical to their biological activity. Both properties are controlled by the manufacture parameters used for the assembly process. Yet little is understood about the evolution of the nanostructure within these complex nanoparticles during assembly, which influences their final form and biological efficacy. Hence in this proposal we aim to use neutron scattering via SANS2D to assess the evolution of LNP nanostructure in microfluidic assembly. We will collect the scattering pattern over different distances away from the mixing junction on a quartz microfluidic mixer. Experiments will be conducted under a range of contrast matching regimes to evaluate the assembly dynamics of the mRNA, lipids and total material independently. The outcomes of this experiment will establish a novel protocol to assess LNP assembly and future design of LNP chemistry and manufacture.