Property-structure-function analysis of complex LNPs using integrative biophysical, molecular-, and cell-based assays

The complexity of lipid nanoparticles (LNPs) for delivery of nucleic acids and oligonucleotides presents a challenge for developability. Fit-for-purpose and complementary analytical tools are required to design successfulformulations, and to inform robust production of stable, safe, and efficacious products. Light scattering technologies remain the front-line methods for physicochemical profiling of LNP vectors throughout their design,processing, development, and quality control. Particle size, polydispersity, and zeta potential are among the recognized critical quality attributes (CQAs) of LNP formulations. Analysis of one or multiple of these keycharacteristics, along with differential scanning calorimetry (DSC) to determine thermal stability profiles and higher order structure (HOS), and cell-based assays to assess LNP transfection efficiency are key for informing theassociation between particle properties and functional performance.