Abstract
Developing effective mRNA vaccines poses certain challenges concerning mRNA stability and ability to induce sufficient immune stimulation and requires a specific panel of techniques for production and testing. Here, we describe the production of stabilized mRNA with enhanced immunogenicity, generated using conventional nucleotides only, by introducing changes to the mRNA sequence and by complexation with the nucleotide-binding peptide protamine (RNActive® technology). Methods described here include the synthesis, purification, and protamine complexation of mRNA vaccines as well as a comprehensive panel of in vitro and in vivo methods for evaluation of vaccine quality and immunogenicity.
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Acknowledgements
This work was partly funded by the German Federal Ministry of Education and Research (BMBF, grant number: 031A061A).
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Rauch, S., Lutz, J., Kowalczyk, A., Schlake, T., Heidenreich, R. (2017). RNActive® Technology: Generation and Testing of Stable and Immunogenic mRNA Vaccines. In: Kramps, T., Elbers, K. (eds) RNA Vaccines. Methods in Molecular Biology, vol 1499. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6481-9_5
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DOI: https://doi.org/10.1007/978-1-4939-6481-9_5
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6479-6
Online ISBN: 978-1-4939-6481-9
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