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

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

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

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.

Original language	English
Article number	103087
Journal	STAR Protocols
Volume	5
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2024.103087
State	Published - 21 Jun 2024

Keywords

biotechnology and bioengineering
cancer
immunology

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10.1016/j.xpro.2024.103087

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title = "Protocol for the development of mRNA lipid nanoparticle vaccines and analysis of immunization efficiency in mice",

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",

author = "Neha Karekar and {Reid Cahn}, Ashley and Judit Morla-Folch and Alexis Saffon and Ward, {Ross W.} and Aparna Ananthanarayanan and Teunissen, {Abraham J.P.} and Nina Bhardwaj and Nicolas Vabret",

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T1 - Protocol for the development of mRNA lipid nanoparticle vaccines and analysis of immunization efficiency in mice

AU - Karekar, Neha

AU - Reid Cahn, Ashley

AU - Morla-Folch, Judit

AU - Saffon, Alexis

AU - Ward, Ross W.

AU - Ananthanarayanan, Aparna

AU - Teunissen, Abraham J.P.

AU - Bhardwaj, Nina

AU - Vabret, Nicolas

PY - 2024/6/21

Y1 - 2024/6/21

N2 - 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.

AB - 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.

KW - biotechnology and bioengineering

KW - cancer

KW - immunology

UR - http://www.scopus.com/inward/record.url?scp=85193774820&partnerID=8YFLogxK

U2 - 10.1016/j.xpro.2024.103087

DO - 10.1016/j.xpro.2024.103087

M3 - Article

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SN - 2666-1667

VL - 5

JO - STAR Protocols

JF - STAR Protocols

IS - 2

M1 - 103087

ER -

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

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Keywords

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