CWA 18386:2026

This document specifies a standardized analytical protocol for the quantification of RNA and DNA encapsulation efficiency in lipid nanoparticles (LNPs) using a fluorometric method. The method is based on the use of fluorescence dyes that selectively interact with free nucleic acids, enabling a clear distinction between encapsulated and non-encapsulated species. To determine the total nucleic acid content, the LNPs are lysed using a detergent-based treatment, allowing complete release of the encapsulated material for accurate measurement.
The protocol described herein is designed to offer a rapid, sensitive, and reproducible approach for the non-destructive quantification of nucleic acid encapsulation efficiency. It addresses the limitations of traditional techniques such as gel electrophoresis and UV spectroscopy, which may lack sensitivity and specificity.
This document is applicable to the characterization of LNP formulations used for nucleic acid delivery, particularly in research and early development phases. It is intended for use by academic laboratories, biotechnology companies, and pharmaceutical developers working on LNP-based systems for therapeutic applications.
The scope of this document is limited to the analytical determination of encapsulation efficiency by fluorometry. It does not cover aspects related to LNP formulation, physicochemical characterization, in vivo performance, or therapeutic efficacy.