1.1 Lipid nanoparticles (LNPs) have emerged as an important nanoscale drug delivery technology for the in vivo intracellular delivery of therapeutic proteins and nucleic acids. The safe and efficient delivery of therapeutic proteins and nucleic acids to cells and tissues can enable the effective treatment and prevention of a wide range of human diseases (for example, cancers, infectious diseases, etc.).

1.2 This guide covers the key technical considerations and describes analytical methodology for establishing high quality characterization of ribonucleic acids (RNAs), specifically messenger RNA (mRNA) encapsulated in LNP drug product formulations (Fig. 1).

1.3 Current industry best practices including suitable analytical methods and tools, (1) for measuring the mRNA encapsulation efficiency (that is, the percent ratio of encapsulated mRNA to total mRNA [free + encapsulated] in LNP nucleic acid delivery vehicles), (2) for the qualitative and/or quantitative estimation of the extraction of LNP encapsulated mRNA and (3) for characterizing the identity, integrity, and purity of mRNA before and after encapsulation in LNP delivery vehicles are all described within this guide.

1.4 The technical guidance in this standard may also have direct applicability to the characterization of self-amplifying RNAs (saRNAs), short/small interfering RNAs (siRNAs), as well as to other nucleic acid drug substances encapsulated into LNP vehicles.

1.5 The scope of this guide is limited to the characterization of mRNA payloads and does not describe the physical characterization of the LNP nor methods to assess in vitro or in vivo potency of the drug product.

1.6 Units—The values stated in SI units are to be regarded as the standard. No other units of measurement are included in this standard.

1.7 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.

1.8 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.