Beyond Dye-Based Encapsulation Assays: Comprehensive Physicochemical Characterization Strategies for LNP-Based Therapeutics

Mateusz Imiolek, Waters, MA, USA
Abraham Finny, Waters, MA, USA
Bala Addepalli, Waters, MA, USA
Steve Koza, Waters, MA, USA
Ying Qing Yu, Waters, MA, USA
Michelle Chen , Waters, MA, USA

The rapid expansion of lipid nanoparticle (LNP) therapeutics has created an urgent need for characterization strategies that move beyond encapsulation efficiency to deliver potency-indicating insights. While applications such as in vivo CAR-T illustrate the promise of gene insertion directly into circulating immune cells, they also underscore the analytical challenges posed by increasingly complex drug substances that integrate nucleic acids, lipids, and protein conjugates.

Our studies on model systems and prototype formulations are designed to address these challenges by developing platform analytical methods that can be broadly applied across therapeutic modalities. Intact-particle analyses—including DLS, ELS, FFF, SEC, and CDMS—provide orthogonal measures of particle size, charge, and mass distributions. Complementary deformulation workflows with liquid chromatography (HILIC, RP, and SEC) enable quantification of encapsulation efficiency and lipid composition, with case studies such as phenyl-hexyl stationary phases highlighting the potential for improved lipid resolution.

Taken together, these strategies establish a foundation for platform characterization methods capable of supporting LNP development across diverse applications—from gene insertion and mRNA replacement to RNAi therapeutics. By embracing both the rising complexity of drug substances and the remarkable progress of genetic medicine, these approaches aim to provide analytical frameworks that ensure potency, quality, and regulatory confidence.

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Short Biography of Presenting Author

Mateusz is a Principal Scientist at Waters, specializing in analytical challenges related to the characterization of emerging biotherapeutic modalities. Working in close collaboration with the Institute of Pharmaceutical Sciences of Western Switzerland, he advances application sciences with a recent focus on chromatographic separations of nucleic acids and lipid nanoparticles (LNPs). He has co-authored more than 15 publications on analytical technologies and methods, frequently partnering with leading industry players to address unmet needs in biomolecule analysis.