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Protocol for the development of mRNA lipid nanoparticle vaccines and analysis of immunization efficiency in mice
Neha Karekar1, Ashley Reid Cahn2, Judit Morla-Folch3
1Department of Immunology and Immunotherapy, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Department of Hematology/Oncology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Abstract:
Here, we present a protocol for the development of mRNA-loaded lipid nanoparticle (LNP) vaccines for target antigen sequences of interest. We describe key steps required to design and synthesize mRNA constructs, their LNP encapsulation, and mouse immunization. We then detail quality control assays to determine RNA purity, guidelines to measure RNA immunogenicity using in vitro reporter systems, and a technique to evaluate antigen-specific T cell responses following immunization.
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