PharmSci360

Presentation abstract:

Lipid nanoparticles (LNPs) are pivotal in nucleic acid delivery technologies, advancing therapeutic applications. However, challenges in manufacturing and characterization must be addressed, including ensuring drug product integrity, achieving sufficient recovery during purification while maintaining functionality, and producing a well-purified product. To achieve precise separation and a uniform, functional product, purification using CIM monolithic columns have been developed. Purification and characterization become increasingly critical as product complexity grows, such as encapsulating multiple payloads or adding targeting moieties to the surface.Monolithic columns can be used for efficient purification of LNPs, as an alternative to standard processes (TFF and dialysis). Mobile phases for purification are optimized for high recovery, stability, and functionality. Compared to standard processes, the novel chromatographic method using CIM monoliths demonstrates superior activity and uniformity due to reduced size distribution and enhanced activity. Integrating monolithic technology ensures consistent product quality and process efficiency, supporting safe and effective LNP-based therapies.

Presentation abstract:

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, 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.