Protocol for the development of mRNA lipid nanoparticle vaccines and analysis of immunization efficiency in mice
Neha Karekar, Ashley Reid, Judit Morlà‐Folch, Alexis Saffon, Ross Ward, Aparna Ananthanarayanan, Abraham J. P. Teunissen, Nina Bhardwaj, Nicolas Vabret
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.
Topics & Concepts
ImmunizationProtocol (science)VirologyChemistryBiologyImmunologyMedicineAntibodyPathologyAlternative medicineRNA Interference and Gene DeliveryImmunotherapy and Immune ResponsesAnimal Virus Infections Studies