Sequence-Optimized mRNA Vaccines Against Infectious Disease

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2786; p. 183
• Main Authors: Rauch, Susanne, Lutz, Johannes, Mühe, Janine, Kowalczyk, Aleksandra, Schlake, Thomas, Heidenreich, Regina
• Format: Journal Article
• Language: English
• Published: United States 2024
• Subjects: Animals; Humans; Immunogenicity, Vaccine; Liposomes; Mice; mRNA Vaccines; Nanoparticles - chemistry; RNA Stability; RNA, Messenger - genetics; RNA, Messenger - immunology; SARS-CoV-2 - genetics; SARS-CoV-2 - immunology; Vaccines, Synthetic - genetics; Vaccines, Synthetic - immunology; Stabilized mRNA; RNActive; mRNA vaccines; GC enrichment; Adjuvanticity
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-3770-8_8

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 vaccines (RNActive
technology) with enhanced immunogenicity, generated using conventional nucleotides only, by introducing changes to the mRNA sequence and by formulation into lipid nanoparticles. Methods described here include the synthesis, purification, and formulation of mRNA vaccines as well as a comprehensive panel of in vitro and in vivo methods for evaluation of vaccine quality and immunogenicity.