Production and Evaluation of Nucleoside-Modified mRNA Vaccines for Infectious Diseases

May 30, 2024Methods in molecular biology (Clifton, N.J.)

Creating and Testing Modified mRNA Vaccines for Infectious Diseases

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Abstract

Lipid nanoparticle-encapsulated mRNA vaccines have shown effectiveness in preclinical models against various pathogens.

Nucleoside modification in mRNA vaccines may enhance safety and immune response in humans.
Incorporating modified nucleosides and purifying mRNA after transcription may reduce inflammation and increase translatability.
Lipid nanoparticles protect mRNA from degradation and promote efficient delivery, leading to sustained protein production.
Certain lipid nanoparticles with ionizable cationic lipids may enhance immune responses when used with mRNA vaccines.

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Lipid nanoparticle (LNP)-encapsulated nucleoside-modified mRNA vaccines have demonstrated potency in multiple preclinical models against various pathogens and have recently received considerable attention due to the success of the two safe and effective COVID-19 mRNA vaccines developed by Moderna and Pfizer-BioNTech. The use of nucleoside modification in mRNA vaccines seems to be critical to achieve a sufficient level of safety and immunogenicity in humans, as illustrated by the results of clinical trials using either nucleoside-modified or unmodified mRNA-based vaccine platforms. It is well documented that the incorporation of modified nucleosides in the mRNA and stringent mRNA purification after in vitro transcription render it less inflammatory and highly translatable; these two features are likely key for mRNA vaccine safety and potency. Formulation of the mRNA into LNPs is important because LNPs protect mRNA from rapid degradation, enabling efficient delivery and high levels of protein production for extended periods of time. Additionally, recent studies have provided evidence that certain LNPs with ionizable cationic lipids (iLNPs) possess adjuvant activity that fosters the induction of strong humoral and cellular immune responses by mRNA-iLNP vaccines.In this chapter we describe the production of iLNP-encapsulated, nucleoside-modified, and purified mRNA and the evaluation of antigen-specific T cell and antibody responses elicited by this vaccine form.

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Production and Evaluation of Nucleoside-Modified mRNA Vaccines for Infectious Diseases

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