Celery root-plant derived vesicles: comprehensive isolation, characterization and proteomic analysis

Despite its potent antioxidant and anti-inflammatory capacities, celery (Apium graveolens) root has not been studied as much as its leaf portion. Plant-derived vesicles (PDVs) are emerging biotechnological tools as biocompatible nanocarriers; however, detailed characterization of celery root-derived PDVs (CR-PDVs) is lacking in the literature. This study aimed to comprehensively characterize the physical and proteomic properties of CR-PDVs for the first time, demonstrating their innovative potential in disease modeling and therapeutic applications.

CR-PDVs were purified using the Aqueous Two-Phase System (ATPS) protocol. Morphology, size distribution, and particle density were measured by Nanoparticle Tracking Analysis (NTA). The vesicle proteome was characterized by LC-MS/MS, and proteins were functionally classified using Gene Ontology and KEGG enrichment analyses. CR-PDVs were homogeneous spherical structures with a modal diameter of 147.6 ± 3 nm and yielded 1.15 × 10¹⁰±10⁸ nanoparticle/mL. A total of 168 proteins were identified; proteomic functional classification showed that the majority of proteins support folding and chaperoning processes, while the remainder clustered into subcategories such as vesicle biogenesis-transport and oxidative stress response. KEGG enrichment analysis revealed that proteasome, ribosome, and phagosome pathways were prominent, suggesting that CR-PDVs may play a functional role in protein synthesis, degradation, and cellular uptake processes.

CR-PDVs, with their high structural integrity and biologically rich cargo, can be evaluated as innovative drug carriers or biomarker platforms in cancer, neurodegenerative, and autoimmune disease models. This study provides the first comprehensive dataset on CR-PDVs, providing a solid foundation for translational research.