Mapping Nanoscale Protein-Corona Kinetics of DoE-Optimized Perfluorocarbon Encapsulated-PLGA Nanoparticles by In...

Integrated frameworks that couple statistically guided formulation with nanoscale, time-resolved mapping of the protein corona (PC) remain rare for perfluoro-15-crown-5-ether-loaded poly(lactic-co-glycolic acid) nanoparticles (PFCE-PLGA NPs). We apply a two-phase workflow: a single-factor screen of PLGA/surfactant molecular-weight (Mw) pairings under homogenization versus probe sonication; and a central composite design varying sonication time, surfactant concentration, and PFCE volume to model effects on size, polydispersity index), zeta (ζ)-potential, loading capacity (LC), and fluorine signal-to-noise ratio. Scanning electron microscopy and transmission electron microscopy confirm spherical morphology of the NPs. Synchrotron small-angle X-ray scattering (SAXS) first resolves PFCE-dependent internal NP structure and then, in situ and time-resolved, tracks corona evolution across physiologic albumin levels (40, 20, and 2 mg mL^-1), quantifying shell thickness, protein volume fraction, and polydispersity while distinguishing PFCE-PLGA from blank PLGA NPs. Fibroblast assays show no significant cytotoxicity at tested doses. Overall, combining design-of-experiments with in situ SAXS links processing variables to nanoscale structure and protein-mediated transformations, guiding rational PFCE-PLGA NP design.