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Updated: Jan 16, 2026

Formulating and Characterizing Lipid Nanoparticles for Gene Delivery using a Microfluidic Mixing Platform
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Capillary-Based Physicochemical Characterization of Lipid Nanoparticles.

Evrim Ümit Kuzucu1, Valentin Schittny1, Jörg Huwyler1

  • 1Division of Pharmaceutical Technology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland.

Electrophoresis
|September 28, 2025
PubMed
Summary

New capillary electrophoresis methods characterize lipid nanoparticles (LNPs) for nucleic acid delivery. This advanced analysis reveals LNP structure and identifies unencapsulated DNA, improving vaccine and gene therapy development.

Keywords:
capillary zone electrophoresis (CZE) | critical quality attributes | electrohydrodynamic coupling | electrophoretic Taylor dispersion (eTD) | gene therapy | lipid nanoparticles | nucleic acids | Taylor dispersion (TD)

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Area of Science:

  • Analytical Chemistry
  • Nanotechnology
  • Biochemistry

Background:

  • Lipid nanoparticles (LNPs) are crucial for delivering nucleic acids (NAs) in gene therapy and mRNA vaccines.
  • Current analytical methods struggle to fully characterize the complex physicochemical properties of LNPs.
  • Accurate characterization of LNPs is essential for ensuring the efficacy and safety of NA-based therapeutics.

Purpose of the Study:

  • To introduce a novel analytical strategy for characterizing deoxyribonucleic acid (DNA)-loaded LNPs.
  • To utilize capillary zone electrophoresis (CZE) and Taylor dispersion (TD) analysis, including a combined electrohydrodynamic coupling (eTD) mode.
  • To assess critical quality attributes (CQAs) of LNPs and differentiate between encapsulated and unencapsulated nucleic acids.

Main Methods:

  • Development and application of a novel electrohydrodynamic coupling (eTD) separation mode using standard capillary electrophoresis (CE) instrumentation.
  • Utilized pressure-driven Taylor dispersion (TD) analysis to determine LNP hydrodynamic radius and NA distribution.
  • Employed capillary zone electrophoresis (CZE) to estimate ζ-potential and DNA localization within particle populations.

Main Results:

  • The eTD mode provided deeper insights into LNP structure and morphology, generating characteristic profiles for different formulations.
  • TD analysis revealed the hydrodynamic radius of LNPs and the distribution of NAs in various chemical environments.
  • CZE and eTD successfully distinguished between encapsulated and unencapsulated single-stranded mRNA and double-stranded DNA, identifying free NAs.

Conclusions:

  • Capillary techniques, including the novel eTD mode, offer advanced physicochemical characterization of nucleic acid-loaded lipid nanoparticles.
  • The developed method accurately assesses critical quality attributes and differentiates encapsulated from free nucleic acids.
  • These findings highlight the potential of capillary methods for improving the analysis and development of LNP-based therapeutics.