Titlebook: RNA Vaccines; Methods and Protocol Thomas Kramps Book 2024Latest edition The Editor(s) (if applicable) and The Author(s), under exclusive l

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Self-Replicating RNA Derived from the Genomes of Positive-Strand RNA Virusesd approaches to novel safe and effective vaccines. The following chapter summarizes the principles how such RNAs can be established and used for design of vaccines. Due to the large variety of strategies needed to circumvent specific pitfalls in the design of such constructs the technical details of

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Generation and Characterization of In Vitro Transcribed mRNA. We then describe methods used for mRNA characterization, including UV and fluorescence spectrophotometry, as well as gel electrophoresis. Moreover, characterization of the in vitro transcribed RNA using the Bioanalyzer instrument is described, allowing a higher resolution analysis of the target mo

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Sequence-Optimized mRNA Vaccines Against Infectious Disease 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 nanop

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Electroporation of mRNA as a Universal Technology Platform to Transfect a Variety of Primary Cells wtypes. We have spent more than two decades to optimize and adapt this method, first for antigen-loading of dendritic cells (DCs) and subsequently for T cells, B cells, bulk PBMCs, and several cell lines. In this regard, antigens were introduced, processed, and presented in context of MHC class I and

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A Basic Method for Formulating mRNA-Lipid Nanoparticle Vaccines in the Labn in 2018, and of the two mRNA SARS-CoV-2 vaccines, BNT162b2 and mRNA-1273 in 2021. A key to the success of these therapeutics lies in the lipid-based delivery system. The therapeutic RNAs are encapsulated in lipid nanoparticles (LNPs), which protect against enzymatic degradation and efficiently del