A rapid and reliable strategy for identification of LNP and protein corona in vitro and in vivo - PubMed
6 hours ago
- #Analytical Chemistry
- #Lipid Nanoparticles
- #Protein Corona
- Developed an integrated analytical method for visual verification and analysis of lipid nanoparticles (LNPs) and protein coronas in vitro and in vivo.
- Used SDS-PAGE with Coomassie brilliant blue staining for rapid visual verification and semi-quantification of LNPs/lipids, achieving a linear standard curve with R² = 0.992.
- Applied LC-MS/MS and GC-MS for specific detection and quantification of key lipids (SM-102, DSPC, DMG-PEG2000, cholesterol), all showing excellent linearity (R² > 0.90).
- Separated LNPs and protein coronas using SDS-PAGE after isolation via size-exclusion chromatography (SEC) or sucrose density gradient centrifugation (S-DGC), with visualization by CBB staining.
- Confirmed intact LNPs in SEC and S-DGC fractions using TEM, and found highly consistent protein coronas enriched in apolipoproteins and immune-related proteins via protein MS.
- Enriched pathways included complement/coagulation and cholesterol metabolism, with in vivo analysis revealing stark compositional differences (e.g., enriched immunoglobulins) compared to in vitro.
- Demonstrated that SDS-PAGE/CBB can indirectly assess LNP uptake by cells, validating a rapid, reliable method for simultaneous LNP and protein corona detection with visual verification to improve quality control.