Protocol for the development of mRNA lipid nanoparticle vaccines and analysis of immunization efficiency in mice

Neha Karekar; Ashley Reid Cahn; Judit Morla-Folch; Alexis Saffon; Ross W Ward; Aparna Ananthanarayanan; Abraham J P Teunissen; Nina Bhardwaj; Nicolas Vabret

doi:10.1016/j.xpro.2024.103087

Protocol for the development of mRNA lipid nanoparticle vaccines and analysis of immunization efficiency in mice

STAR Protoc. 2024 Jun 21;5(2):103087. doi: 10.1016/j.xpro.2024.103087. Epub 2024 May 23.

Authors

Neha Karekar¹, Ashley Reid Cahn², Judit Morla-Folch³, Alexis Saffon⁴, Ross W Ward⁵, Aparna Ananthanarayanan⁵, Abraham J P Teunissen³, Nina Bhardwaj⁵, Nicolas Vabret⁶

Affiliations

¹ Department 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.
² Department 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; Icahn Genomics Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
³ Biomedical Engineering and Imaging Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Cardiovascular Research Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA; Icahn Genomics Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁴ Department 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; Icahn Genomics Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁵ 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.
⁶ Department 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; Icahn Genomics Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. Electronic address: [email protected].

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.

Keywords: biotechnology and bioengineering; cancer; immunology.

MeSH terms

Animals
Immunization* / methods
Lipids* / chemistry
Liposomes
Mice
Nanoparticles* / chemistry
Nanovaccines
RNA, Messenger* / genetics
Vaccines / administration & dosage
Vaccines / chemistry
Vaccines / immunology
mRNA Vaccines / immunology

Substances

RNA, Messenger
Lipids
Lipid Nanoparticles
Vaccines
mRNA Vaccines
Nanovaccines
Liposomes