A novel mechanism for the loss of mRNA activity in lipid nanoparticle delivery systems

Nat Commun. 2021 Nov 22;12(1):6777. doi: 10.1038/s41467-021-26926-0.


Lipid nanoparticle (LNP)-formulated mRNA vaccines were rapidly developed and deployed in response to the SARS-CoV-2 pandemic. Due to the labile nature of mRNA, identifying impurities that could affect product stability and efficacy is crucial to the long-term use of nucleic-acid based medicines. Herein, reversed-phase ion pair high performance liquid chromatography (RP-IP HPLC) was used to identify a class of impurity formed through lipid:mRNA reactions; such reactions are typically undetectable by traditional mRNA purity analytical techniques. The identified modifications render the mRNA untranslatable, leading to loss of protein expression. Specifically, electrophilic impurities derived from the ionizable cationic lipid component are shown to be responsible. Mechanisms implicated in the formation of reactive species include oxidation and subsequent hydrolysis of the tertiary amine. It thus remains critical to ensure robust analytical methods and stringent manufacturing control to ensure mRNA stability and high activity in LNP delivery systems.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehydes / chemistry
  • Chromatography, Liquid
  • Drug Delivery Systems*
  • Humans
  • Ions / chemistry
  • Lipids / chemistry
  • Liposomes / chemistry*
  • Nanoparticles / chemistry*
  • Nucleosides / chemistry
  • Oxidation-Reduction
  • Protein Biosynthesis
  • RNA Stability
  • RNA, Messenger / chemistry*
  • Vaccine Potency*
  • mRNA Vaccines / chemistry


  • Aldehydes
  • Ions
  • Lipid Nanoparticles
  • Lipids
  • Liposomes
  • Nucleosides
  • RNA, Messenger
  • mRNA Vaccines