Resolving Heterogeneity of Targeted Lipid Nanoparticles Through Solution-Based Biophysical Analyses.

Authors

Hannah C Geisler, Hannah C Safford, Ajay S Thatte, Marshall S Padilla, Elisa Battistini, Hannah M Yamagata, Violet M Ullman, Alex Chan, Benjamin E Nachod, Anushka Agrawal, Maxwell B Watkins, Jesse B Hopkins, Andrew Tsourkas, Kushol Gupta, Michael J Mitchell

Published in

Advanced materials (Deerfield Beach, Fla.). Pages e73984. Jul 21, 2026. Epub Jul 21, 2026.

Abstract

Targeted lipid nanoparticles (tLNPs) enable cell-specific nucleic acid delivery through covalent attachment of targeting ligands that drive receptor-mediated LNP uptake. tLNPs are potentially promising for pregnancy‑associated applications where precise delivery is required to minimize maternal toxicity and protect fetal health. Yet, rational tLNP design is limited by an incomplete understanding of how physicochemical properties influence biological performance. Traditional analytical methods report only ensemble-averaged properties, leaving the nanoscale heterogeneity of tLNPs unresolved. Here, we utilize asymmetric flow field-flow fractionation integrated with in-line UV spectral analysis, light scattering, and synchrotron small-angle X-ray scattering (AF4-UV-DLS-MALS-SAXS) to resolve ligand-dependent tLNP subpopulations that differ in size, shape, composition, and relative abundance. Protein conjugation preserves the internal lipid-RNA nanostructure of base LNPs but substantially increases particle heterogeneity, particularly for larger and multivalent ligands. Despite increased heterogeneity, tLNPs functionalized with higher-avidity ligands achieve more effective targeted placental RNA delivery in mice. Chemometric SAXS analyses reveal that only SAXS-resolved tLNP subpopulations, not ensemble-averaged parameters, correlate with targeted placental transfection in vivo, whereas bulk physicochemical metrics more strongly associate with nonspecific hepatic delivery. Together, this work harnesses a separation-coupled biophysical platform to resolve previously inaccessible tLNP subpopulations and provides insights to inform rational engineering of next-generation targeted RNA therapeutics.

PMID:
42478324
Bibliographic data and abstract were imported from PubMed on 21 Jul 2026.

Read full publication at:
Please sign in to see all details.

Stats

1-terrible, 9-excellent. How would you rate this publication? Sign in in to submit your rating.

Post a comment

You need to be signed in to post comments. You can sign in here.

Comments

There are no comments yet.