Mapping Nanoscale Protein‐Corona Kinetics of DoE‐Optimized Perfluorocarbon Encapsulated‐PLGA Nanoparticles by In Situ, Time‐Resolved Synchrotron SAXS

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ABSTRACT 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.

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