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Our method relies upon established laboratory techniques, can be tailored to a variety of biological questions regarding the pharmacokinetic disposition of extracellular vesicles, and will provide a complementary approach for the of study EV ligand-receptor interactions in the context of EV uptake and targeted therapeutics.Įxtracellular vesicles (EV) can be used to improve medical treatments if properly understood ( 1, 2). Covariate and bootstrap analyses identified cell type having an influence on peripheral volumes (V2 and V3) and clearance (Cl3). We performed an observation-based simulated posterior predictive evaluation with prediction-corrected visual predictive check. Data were best described by a three-compartment model with one elimination from the central compartment. Incorporated oligonucleotide was stable in blood and detectable over five half-lives. ResultsĨ6.5% ± 1.5% (mean ± S.E.) of EV particles were in the 45–195 nm size range and demonstrated protein and lipid markers of endosomal origin. Non-linear mixed effects analysis with first order conditional estimation – extended least squares (FOCE ELS) was used to estimate population-level parameters with associated intra-animal variability. Digital PCR was leveraged to allow for quantification over a wide dynamic range. Jugular vein catheters were used to introduce EVs to conscious rats (n = 30) and to collect blood samples. MethodsĬrude sEVs were labeled with a non-homologous oligonucleotide and isolated from cell culture media using a commercial reagent. Using labeled sEVs administered to conscious rats, we developed a multiple compartment pharmacokinetic model to identify potential differences in the disposition of sEVs from three different cell types.
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We developed an accessible method for labeling small extracellular vesicles (sEVs) without disrupting endogenous ligands.
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