Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Feb 7;12(2):169.
doi: 10.3390/vaccines12020169.

Correlating Stability-Indicating Biochemical and Biophysical Characteristics with In Vitro Cell Potency in mRNA LNP Vaccine

Xin Tong  1 , Jessica Raffaele  1 , Katrina Feller  1 , Geethanjali Dornadula  1 , James Devlin  1 , David Boyd  2 , John W Loughney  1 , Jon Shanter  2 , Richard R Rustandi  1
Affiliations

Correlating Stability-Indicating Biochemical and Biophysical Characteristics with In Vitro Cell Potency in mRNA LNP Vaccine

Xin Tong et al. Vaccines (Basel). .

Abstract

The development of mRNA vaccines has increased rapidly since the COVID-19 pandemic. As one of the critical attributes, understanding mRNA lipid nanoparticle (LNP) stability is critical in the vaccine product development. However, the correlation between LNPs' physiochemical characteristics and their potency still remains unclear. The lack of regulatory guidance on the specifications for mRNA LNPs is also partially due to this underexplored relationship. In this study, we performed a three-month stability study of heat-stressed mRNA LNP samples. The mRNA LNP samples were analyzed for their mRNA degradation, LNP particle sizes, and mRNA encapsulation efficiency. In vitro cell potency was also evaluated and correlated with these above-mentioned physiochemical characterizations. The mRNA degradation-cell potency correlation data showed two distinct regions, indicating a critical cut-off size limit for mRNA degradation. The same temperature dependence was also observed in the LNP size-cell potency correlation.

Keywords: lipid nanoparticles; mRNA vaccines; potency; stability.

PubMed Disclaimer

Conflict of interest statement

X.T., J.R., K.F., G.D., J.D., D.B., J.W.L., J.S. and R.R.R. are/were all employees of Merck Sharp & Dohme LLC, a subsidiary of Merck & Co., Inc., Rahway, NJ, USA, and may hold stock in Merck & Co., Inc., Rahway, NJ, USA.

Figures

Figure 1
Figure 1
(A) mRNA integrity (%intact mRNA, y-axis) was plotted against time (days, x-axis). (B) Cell-based relative potency (y-axis) was plotted against time (days, x-axis). (C) Correlations between %intact mRNA (integrity) in the y-axis and the cell-based relative potency in the x-axis. Two colored circles (yellow and red) indicate two distinguished subsets. Two colored solid lines (yellow and purple) indicate positive and zero correlations, respectively. Each data point represents the average result tested at a single time point within their respective storage condition. For in vitro potency and mRNA integrity testing, samples were tested in duplicate (n = 2).
Figure 2
Figure 2
(A) Correlations between the LNP particle size in the y-axis and the cell-based relative potency in the x-axis with all the data points. (B) Correlations between the LNP particle size and the cell-based relative potency with only frozen samples (−20 °C and −70 °C). Each data point represents the average result tested at a single time point within its respective storage condition. For in vitro potency and particle size testing, samples were tested in duplicate (n = 2).
Figure 3
Figure 3
Correlations between the LNP encapsulation efficiency (%EE) in the y-axis and the cell-based relative potency in the x-axis. Each data point represents the average result tested at a single time point within its respective storage condition. For in vitro potency testing, samples were tested in duplicate (n = 2). For encapsulation efficiency testing, samples were tested in quadruplicate (n = 4).
See this image and copyright information in PMC

References

    1. Lorentzen C.L., Haanen J.B., Met O., Svane I.M. Clinical advances and ongoing trials on mRNA vaccines for cancer treatment. Lancet Oncol. 2022;23:e450–e458. doi: 10.1016/S1470-2045(22)00372-2. - DOI - PMC - PubMed
    1. Park J.W., Lagniton P.N.P., Liu Y., Xu R.H. mRNA vaccines for COVID-19: What, why and how. Int. J. Biol. Sci. 2021;17:1446–1460. doi: 10.7150/ijbs.59233. - DOI - PMC - PubMed
    1. Geall A.J., Verma A., Otten G.R., Shaw C.A., Hekele A., Banerjee K., Cu Y., Beard C.W., Brito L.A., Krucker T., et al. Nonviral delivery of self-amplifying RNA vaccines. Proc. Natl. Acad. Sci. USA. 2012;109:14604–14609. doi: 10.1073/pnas.1209367109. - DOI - PMC - PubMed
    1. Pardi N., Tuyishime S., Muramatsu H., Kariko K., Mui B.L., Tam Y.K., Madden T.D., Hope M.J., Weissman D. Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to mice by various routes. J. Control. Release. 2015;217:345–351. doi: 10.1016/j.jconrel.2015.08.007. - DOI - PMC - PubMed
    1. Zhang C.L., Maruggi G., Shan H., Li J.W. Advances in mRNA Vaccines for Infectious Diseases. Front. Immunol. 2019;10:594. doi: 10.3389/fimmu.2019.00594. - DOI - PMC - PubMed

LinkOut - more resources