Insights into mRNA Lipid Nanoparticle Polydispersity and Shape of Using Quantitative Solution Biophysics

Lipid nanoparticles (LNPs) are crucial for RNA therapeutics, notably after the success of Patisiran and COVID-19 mRNA vaccines. LNPs need precise design for effective clinical use, but the understanding on mixing methods influences the LNP structure and biological activity is limited.

This study addresses this gap by comparing standard and advanced characterization methods including multi-wavelength analytical ultracentrifugation (MWL-AUC), multi-angle light scattering (MALS), and synchrotron size-exclusion chromatography small-angle X-ray scattering (SEC-SAXS) with singular-value decomposition methods (SVD). We will present the strengths and weaknesses of each approach and showcase the increased detail newer advanced methods provide by comparing LNP formulations made using two common small-scale production methods: microfluidic rapid mixing and bulk mixing. The characterization techniques employed here can enhance our understanding of LNP structure-function relationships and enable researchers to define their RNA LNP products more precisely, which can improve LNP quality and potentially accelerate pharmaceutical development.