Using Orthogonal Analytical Approaches to Profile Lipid Nanoparticle Physicochemical Attributes

Abstract Lipid nanoparticles (LNPs) have become a major disruptor within the drug delivery field of complex RNA molecules. The wide applicability of prototype nanomedicines has the potential to fill clinical requirements for use against current untreatable diseases. The uptake and implementation of analytical technologies to evaluate these prototype nanomedicines have not experienced similar growth rates, thus hindering the translation of LNPs. Here, we evaluate a model RNA-LNP formulation with a selection of routine and high-resolution orthogonal analytical techniques across studies on the manufacturing process parameter impact and formulation stability evaluation under refrigerated and ultra-low temperatures. We analysed a model cationic RNA-complexed LNP formulation via the process impact on formulation critical quality attributes, short-term refrigerated stability evaluation and frozen-storage stability using zetasizer dynamic light scattering and nanoparticle tracking analysis. We also evaluated freeze-/thaw-induced stress on LNP formulation using high-resolution field-flow fractionation. Statistical analysis and correlations between techniques were conducted to further enhance our understanding of LNP formulation design and its physicochemical attributes to facilitate LNP formulation clinical translation.