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Taylor Dispersion Analysis to support lipid-nanoparticle formulations for mRNA vaccines

Gene Therapy volume 30, pages 421–428 (2023)Cite this article

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Abstract

Lipid nanoparticles (LNPs) are currently the most advanced non-viral clinically approved messenger ribonucleic acid (mRNA) delivery systems. The ability of a mRNA vaccine to have a therapeutic effect is related to the capacity of LNPs to deliver the nucleic acid intact into cells. The role of LNPs is to protect mRNA, especially from degradation by ribonucleases (RNases) and to allow it to access the cytoplasm of cells where it can be translated into the protein of interest. LNPs enter cells by endocytosis and their size is a critical parameter impacting their cellular internalization. In this work, we studied different formulation process parameters impacting LNPs size. Taylor dispersion analysis (TDA) was used to determine the LNPs size and size distribution and the results were compared with those obtained by Dynamic Light Scattering (DLS). TDA was also used to study both the degradation of mRNA in the presence of RNases and the percentage of mRNA encapsulation within LNPs.

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Fig. 1: Schematic representation of lipid nanoparticles (LNPs) formulation by rapid dilution of an ethanolic phase containing lipids in an aqueous phase containing mRNA.
Fig. 2: Taylorgrams obtained for the size characterization of LNPs.
Fig. 3: Size distributions obtained by TDA and DLS.
Fig. 4: Taylorgrams obtained in the absence or in the presence of RNase A.
Fig. 5: Taylorgrams obtained for the determination of percentage of mRNA encapsulation and free mRNA hydrodynamic diameters.

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Additional data are available in the Supplementary Information file.

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Funding

This work was partly funded by Sanofi Pasteur under a Cooperative Research and Development Agreement with the University of Montpellier and the CNRS.

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Authors and Affiliations

  1. IBMM, University of Montpellier, CNRS, ENSCM, Montpellier, France

    Camille Malburet, Laurent Leclercq & Hervé Cottet

  2. Sanofi Pasteur, 1541 avenue Marcel Mérieux, 69280, Marcy l’Etoile, France

    Camille Malburet, Jean-François Cotte, Jérôme Thiebaud, Emilie Bazin & Marie Garinot

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  1. Camille Malburet
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Contributions

CM, LL, J-FC, JT, and HC designed this study. CM and EB formulated the LNPs. CM performed the TDA and DLS experiments. LL, J-FC, JT, EB, MG, and HC reviewed the data. CM wrote the manuscript. LL, J-FC, JT, EB, MG, and HC reviewed and edited the manuscript.

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Correspondence to Hervé Cottet.

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Camille Malburet, Jean-François Cotte, Jérôme Thiebaud, Emilie Bazin and Marie Garinot are Sanofi employees and may hold shares or stocks in the company.

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Malburet, C., Leclercq, L., Cotte, JF. et al. Taylor Dispersion Analysis to support lipid-nanoparticle formulations for mRNA vaccines. Gene Ther 30, 421–428 (2023). https://doi.org/10.1038/s41434-022-00370-1

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