Quantitative analysis of mRNA-lipid nanoparticle stability in human plasma and serum by size-exclusion chromatography coupled with dual-angle light scattering

• Published in: Nanomedicine 2024-06, Vol.58, p.102745-102745, Article 102745
• Main Authors: Liau, Brian, Zhang, Li, Ang, Melgious Jin Yan, Ng, Jian Yao, C.V., Suresh Babu, Schneider, Sonja, Gudihal, Ravindra, Bae, Ki Hyun, Yang, Yi Yan
• Format: Article
• Language: English
• Subjects: Dual-angle light scattering; Lipid nanoparticles; mRNA; Size-exclusion chromatography; Stability
• ISSN: 1549-9634; 1549-9642
• DOI: 10.1016/j.nano.2024.102745

Understanding the stability of mRNA loaded lipid nanoparticles (mRNA-LNPs) is imperative for their clinical development. Herein, we propose the use of size-exclusion chromatography coupled with dual-angle light scattering (SEC-MALS) as a new approach to assessing mRNA-LNP stability in pure human serum and plasma. By applying a dual-column configuration to attenuate interference from plasma components, SEC-MALS was able to elucidate the degradation kinetics and physical property changes of mRNA-LNPs, which have not been observed accurately by conventional dynamic light scattering techniques. Interestingly, both serum and plasma had significantly different impacts on the molecular weight and radius of gyration of mRNA-LNPs, suggesting the involvement of clotting factors in desorption of lipids from mRNA-LNPs. We also discovered that a trace impurity (~1 %) in ALC-0315, identified as its O-tert-butyloxycarbonyl-protected form, greatly diminished mRNA-LNP stability in serum. These results demonstrated the potential utility of SEC-MALS for optimization and quality control of LNP formulations. Size-exclusion chromatography coupled with dual-angle light scattering (SEC-MALS) was used as a new methodology for the characterization of mRNA-LNP stability in undiluted human plasma and serum. The degradation process of mRNA-LNPs in plasma followed first-order kinetics with a half-life of around 85 min, accompanied by a displacement of the lipid components by plasma proteins. Surprisingly, trace contaminants (~1%) in the ionizable lipid significantly affected the stability of mRNA-LNPs in serum. [Display omitted] •The first study demonstrating use of SEC-MALS for quantitative stability analysis of mRNA-LNPs in pure human serum and plasma•mRNA-LNPs degrade with first-order kinetics (half-life: 85 min), and lipid components displaced by plasma proteins.•Even trace amounts (~1 %) of impurity in the ionizable lipid can potentially compromise mRNA-LNP stability in serum