Higher or lower? the resolution of analytical pipelines for the evaluation of lipid nanoparticle critical quality attributes

Dynamic light scattering (DLS) is a routinely used analytical technique to measure the size distribution of a colloidal sample. Nanoparticle tracking analysis (NTA) has more recently emerged as an orthogonal technique to determine particle size and estimated concentration, although widespread use of NTA for lipid nanoparticle (LNP) analysis is not reported. Here, we use DLS and NTA to screen the stability of oligonucleotide drug loaded lipid nanoparticles (oligo-LNPs) at ultra-low storage temperatures (-80 °C) in the presence and absence of a cryoprotectant (20 % sucrose, w/v) as a case study to highlight differences in technique resolution. NTA was able to detect additional LNP subpopulations samples for all measured samples, whereas DLS was unable to detect these subpopulations. Our study conveys that the use of orthogonal sizing techniques can support early-stage product development, where a more in-depth analysis of formulation and process parameters on LNP stability would support comprehensive understanding of variable defining product stability.