Successful batch and continuous lyophilization of mRNA LNP formulations depend on cryoprotectants and ionizable lipids
- Author
- Alexander Lamoot, Joris Lammens (UGent) , Emily De Lombaerde (UGent) , Zifu Zhong (UGent) , Mark Gontsarik (UGent) , Yong Chen (UGent) , Thomas De Beer (UGent) and Bruno De Geest (UGent)
- Organization
- Project
- 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.
- Keywords
- General Materials Science, Biomedical Engineering
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-01HMB4G6ZNAG5AYZK447CJ21PM
- MLA
- Lamoot, Alexander, et al. “Successful Batch and Continuous Lyophilization of MRNA LNP Formulations Depend on Cryoprotectants and Ionizable Lipids.” BIOMATERIALS SCIENCE, vol. 11, no. 12, 2023, pp. 4327–34, doi:10.1039/d2bm02031a.
- APA
- Lamoot, A., Lammens, J., De Lombaerde, E., Zhong, Z., Gontsarik, M., Chen, Y., … De Geest, B. (2023). Successful batch and continuous lyophilization of mRNA LNP formulations depend on cryoprotectants and ionizable lipids. BIOMATERIALS SCIENCE, 11(12), 4327–4334. https://doi.org/10.1039/d2bm02031a
- Chicago author-date
- Lamoot, Alexander, Joris Lammens, Emily De Lombaerde, Zifu Zhong, Mark Gontsarik, Yong Chen, Thomas De Beer, and Bruno De Geest. 2023. “Successful Batch and Continuous Lyophilization of MRNA LNP Formulations Depend on Cryoprotectants and Ionizable Lipids.” BIOMATERIALS SCIENCE 11 (12): 4327–34. https://doi.org/10.1039/d2bm02031a.
- Chicago author-date (all authors)
- Lamoot, Alexander, Joris Lammens, Emily De Lombaerde, Zifu Zhong, Mark Gontsarik, Yong Chen, Thomas De Beer, and Bruno De Geest. 2023. “Successful Batch and Continuous Lyophilization of MRNA LNP Formulations Depend on Cryoprotectants and Ionizable Lipids.” BIOMATERIALS SCIENCE 11 (12): 4327–4334. doi:10.1039/d2bm02031a.
- Vancouver
- 1.Lamoot A, Lammens J, De Lombaerde E, Zhong Z, Gontsarik M, Chen Y, et al. Successful batch and continuous lyophilization of mRNA LNP formulations depend on cryoprotectants and ionizable lipids. BIOMATERIALS SCIENCE. 2023;11(12):4327–34.
- IEEE
- [1]A. Lamoot et al., “Successful batch and continuous lyophilization of mRNA LNP formulations depend on cryoprotectants and ionizable lipids,” BIOMATERIALS SCIENCE, vol. 11, no. 12, pp. 4327–4334, 2023.
@article{01HMB4G6ZNAG5AYZK447CJ21PM,
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.}},
author = {{Lamoot, Alexander and Lammens, Joris and De Lombaerde, Emily and Zhong, Zifu and Gontsarik, Mark and Chen, Yong and De Beer, Thomas and De Geest, Bruno}},
issn = {{2047-4830}},
journal = {{BIOMATERIALS SCIENCE}},
keywords = {{General Materials Science,Biomedical Engineering}},
language = {{eng}},
number = {{12}},
pages = {{4327--4334}},
title = {{Successful batch and continuous lyophilization of mRNA LNP formulations depend on cryoprotectants and ionizable lipids}},
url = {{http://doi.org/10.1039/d2bm02031a}},
volume = {{11}},
year = {{2023}},
}
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