DOE OSTI · 3001470
Elucidating lipid nanoparticle properties and structure through biophysical analyses
Abstract
Designing lipid nanoparticle (LNP) delivery systems with specific targeting, potency and minimal side effects is crucial for their clinical use. However, traditional characterization methods, such as dynamic light scattering, cannot accurately quantify physicochemical properties of LNPs and how these are influenced by the lipid composition and mixing method. Here, we structurally characterize polydisperse LNP formulations by applying emerging solution-based biophysical methods that have higher resolution and provide biophysical data beyond size and polydispersity. These techniques include sedimentation velocity analytical ultracentrifugation, field-flow fractionation followed by multiangle light scattering and size-exclusion chromatography in line with synchrotron small-angle X-ray scattering. Here, we show that LNPs have intrinsic polydispersity in size, RNA loading and shape, which depend on both the formulation technique and the lipid composition. Lastly, we predict LNP transfection in vitro and in vivo by examining the relationship between mRNA translation and physicochemical characteristics. Solution-based biophysical methods will be essential for determining LNP structure–function relationships, facilitating the creation of new design rules for LNPs.
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Padilla, Marshall S. [Univ. of Pennsylvania, Philadelphia, PA (United States)] (ORCID:000000033607790X), Shepherd, Sarah J. [Univ. of Pennsylvania, Philadelphia, PA (United States)], Hanna, Andrew R. [Univ. of Pennsylvania, Philadelphia, PA (United States)] (ORCID:0009000932204378), Kurnik, Martin [Waters Corporation, Goleta, CA (United States)] (ORCID:0000000192559730), Zhang, Xujun [Waters Corporation, Goleta, CA (United States)], Chen, Michelle [Waters Corporation, Goleta, CA (United States)], Byrnes, James [Brookhaven National Laboratory (BNL), Upton, NY (United States). National Synchrotron Light Source II (NSLS-II)] (ORCID:0000000249060640), Joseph, Ryann A. [Univ. of Pennsylvania, Philadelphia, PA (United States)] (ORCID:0000000207956094), Yamagata, Hannah M. [Univ. of Pennsylvania, Philadelphia, PA (United States). Dept. of Physics and Astronomy] (ORCID:0000000295257102), Ricciardi, Adele S. [Univ. of Pennsylvania, Philadelphia, PA (United States); Children’s Hospital of Philadelphia, PA (United States)] (ORCID:0000000326378522), Mrksich, Kaitlin [Univ. of Pennsylvania, Philadelphia, PA (United States)] (ORCID:0009000879712129), Issadore, David [Univ. of Pennsylvania, Philadelphia, PA (United States)] (ORCID:0000000254618653), Gupta, Kushol [Univ. of Pennsylvania, Philadelphia, PA (United States)], Mitchell, Michael J. [Univ. of Pennsylvania, Philadelphia, PA (United States)] (ORCID:0000000236282244). 2025-10-23. Elucidating lipid nanoparticle properties and structure through biophysical analyses. https://doi.org/10.1038/s41587-025-02855-x
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