Issue 12, 2023

Successful batch and continuous lyophilization of mRNA LNP formulations depend on cryoprotectants and ionizable lipids

Abstract

The limited thermostability and need for ultracold storage conditions are the major drawbacks of the currently used nucleoside-modified lipid nanoparticle (LNP)-formulated messenger RNA (mRNA) vaccines, which hamper the distribution of these vaccines in low-resource regions. The LNP core contains, besides mRNA and lipids, a large fraction of water. Therefore, encapsulated mRNA, or at least a part of it, is subjected to hydrolysis mechanisms similar to unformulated mRNA in an aqueous solution. It is likely that the hydrolysis of mRNA and colloidal destabilization are critical factors that decrease the biological activity of mRNA LNPs upon storage under ambient conditions. Hence, lyophilization as a drying technique is a logical and appealing method to improve the thermostability of these vaccines. In this study, we demonstrate that mRNA LNP formulations comprising a reduction-sensitive ionizable lipid can be successfully lyophilized, in the presence of 20% w/v sucrose, both by conventional batch freeze-drying and by an innovative continuous spin lyophilization process. While the chemical structure of the ionizable lipid did not affect the colloidal stability of the LNP after lyophilization and redispersion in an aqueous medium, we found that the ability of LNPs to retain the mRNA payload stably encapsulated, and mediate in vivo and in vitro mRNA translation into protein, post lyophilization strongly depended on the ionizable lipid in the LNP formulation.

Graphical abstract: Successful batch and continuous lyophilization of mRNA LNP formulations depend on cryoprotectants and ionizable lipids

Supplementary files

Article information

Article type
Paper
Submitted
09 dec 2022
Accepted
29 mar 2023
First published
04 apr 2023

Biomater. Sci., 2023,11, 4327-4334

Successful batch and continuous lyophilization of mRNA LNP formulations depend on cryoprotectants and ionizable lipids

A. Lamoot, J. Lammens, E. De Lombaerde, Z. Zhong, M. Gontsarik, Y. Chen, T. R. M. De Beer and B. G. De Geest, Biomater. Sci., 2023, 11, 4327 DOI: 10.1039/D2BM02031A

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements