Nejc Pavlin, Mojca Bavčar, Tristan Kovačič, Tjaša Kašček, Anže Martinčič Celjar, Ines Bergoč, Andreja Gramc Livk, and Aleš Štrancar
Journal of Chromatography B, Volume 1265, 2025.
Abstract:
Lipid nanoparticles (LNPs) are highly versatile delivery systems; however, their complex composition, morphology, and instability pose significant analytical challenges. This study presents the PATfix LNP Switcher Platform, an all-in-one multiparameter solution that addresses key quality parameters crucial for quality control and development.
This is a streamlined two-dimensional chromatographic platform featuring two columns. The first, the HIC CIMac OH column (6 μm), separates LNPs from free nucleic acids by capturing LNP particles, while free nucleic acids do not bind. The second is a reverse-phase CIMac SDVB column (2 μm) with an ion-pairing reagent, which separates and quantifies nucleic acids.
The PATfix LNP Switcher directly injects LNP formulations without pre-treatment, measuring encapsulated and non-encapsulated nucleic acids, their integrity, and size. It detects multiple nucleic acids within the same particle, outperforming dye-based assays like RiboGreen, and preserves LNP structure during analysis to prevent artifacts. This is an analytical solution for overcoming CMC and manufacturing challenges of complex LNP-based biopharmaceuticals for cancer, gene editing, and vaccines.
