Next Generation LNP Pharmaceuticals Analytics: Intact Chromatography for Comprehensive Characterization

Lipid nanoparticles (LNPs) provide the most advanced platform for in vivo drug delivery of nucleic acids.
However, transforming from the development of lipid nanoparticle-based therapeutics towards
commercialized biopharmaceuticals is accompanied by multiple challenges. They include biological,
SUBMISSION PREVIEW: RNA-LNP DRUG PRODUCT: LIPID STABILITY, RNA
IMPURITIES AND PARTICLE CHARACTERIZATION
RNA-LNP Drug Product: Lipid Stability, RNA Impurities and Particle Characterization
Submission ID: 1943482
Submission Type: 30-min Scientific Symposium Presentation
Submission Status: Complete / Locked
manufacturing, and characterization challenges. The focus of characterization is on determining drug
substance purity, excipient purity and the entire drug product purity.
Liquid chromatography is the most widely used tool for routine analytics of pharmaceuticals. Its
application to large biomolecules is more challenging and requires columns with low shear stress and
large pores, such as the monolithic columns. Additonal surface modifications of monolithic columns
enable multifaceted characterization of complex biopharmaceuticals.
Characterization of the excipients, i.e. lipids, as well as, RNA drug substances is of crucial importance for
process development and product quality. In particular, the ionizable lipid, with its fully synthesizable
small molecule with no chromophores and high hydrophobicity, is often hard to purify. It is crucial to
control its purity, but it is even more important to control its impact on the RNA-LNP product manifested
by the interaction of impurities with RNA and the proper formation of the LNP and its subsequent
stability. And then comparing the purity of the RNA pre and post encapsulation. An important aspect of
LNP analytical methods is that they do not require labeling and sample preparation, thus being able to
directly quantify the RNA by UV and lipids by CAD/ELSD .
The supramolecular assembly of the RNA with the lipids surrounding forms the final RNA-LNP drug
product. This one is the least stable and the hardest to characterize intact. Parameters that can describe
it are encapsulation efficiency, particle size, heterogeneity, surface charge, morphology and RNA loading.
They mostly require expensive advance biophysical analytics but can be determined by chromatography
as well.
The chromatography methods were developed with the aim to improve the speed and quality of process
underpinning the development of novel RNA-LNPs.