Simple quantification of mRNA–LNP encapsulation efficiency with scatter-free absorption spectroscopy
Aida López Espinar – Field Application Scientist, Marama Labs IE; Kirsty Smith – Head of Applications, Marama Labs IE; Piotr Kowalski – Associate Professor, School of Pharmacy, APC Microbiome Ireland, University College Cork; Brendan Darby – Chief Executive Officer, Marama Labs IE
Professor of Physics Victoria University of Wellington / Marama Labs Wellington, New Zealand
Introduction: Reliable measurement of total RNA and encapsulation efficiency (EE) is essential for RNA–lipid nanoparticle (LNP) development and manufacturing [1]. Conventional fluorescent-dye EE workflows typically require nanoparticle lysis with surfactants, multiple calibrations, and substantial operator handling, contributing to low repeatability. Scatter-Free Absorbance (SFA) spectroscopy [2], a variation of UV/Vis spectroscopy using an integrating sphere, was recently introduced as a new method to quantify total RNA concentration inside LNPs [3,4]. We here show that it can also be used to measure fluorescence and, combined with RibogreenTM, to quantify free-RNA and therefore EE.
Learning Objectives:
Understand scatter-free absorption (SFA) spectroscopy and its application to RNA quantification in LNPs
Understand how total, free RNA, and EE, can be quantified simply by SFA using the same instrument/set-up.
