A Basic Method for Formulating mRNA-Lipid Nanoparticle Vaccines in the Lab

Methods Mol Biol. 2024:2786:237-254. doi: 10.1007/978-1-0716-3770-8_11.

Abstract

During recent years, RNA therapeutics have begun to make a substantial impact in the clinic, with the approval of the siRNA-based therapeutic Patisiran in 2018, and of the two mRNA SARS-CoV-2 vaccines, BNT162b2 and mRNA-1273 in 2021. A key to the success of these therapeutics lies in the lipid-based delivery system. The therapeutic RNAs are encapsulated in lipid nanoparticles (LNPs), which protect against enzymatic degradation and efficiently deliver the RNA across the cell membrane into the cytosol. Thereby, the method used for LNP synthesis and its lipid composition are crucial aspects that decide the efficacy of the LNP-RNA hetero system. Here we provide a detailed guide for the simple preparation of LNP-encapsulated mRNA vaccines.

Keywords: Infectious diseases; Lipid nanoparticles; Messenger RNA; Microfluidic mixing; Modified nucleoside; Pseudo uridine; Unmodified nucleoside; Vaccine.

MeSH terms

  • 2019-nCoV Vaccine mRNA-1273
  • BNT162 Vaccine
  • COVID-19 Vaccines* / immunology
  • COVID-19* / prevention & control
  • COVID-19* / virology
  • Humans
  • Lipids* / chemistry
  • Liposomes / chemistry
  • Nanoparticles* / chemistry
  • Nanovaccines
  • RNA, Messenger* / genetics
  • SARS-CoV-2* / genetics
  • SARS-CoV-2* / immunology
  • mRNA Vaccines

Substances

  • COVID-19 Vaccines
  • Lipids
  • RNA, Messenger
  • Lipid Nanoparticles
  • 2019-nCoV Vaccine mRNA-1273
  • BNT162 Vaccine
  • mRNA Vaccines
  • Liposomes
  • Nanovaccines