Isolation and characterization of ginseng-derived exosome-like nanoparticles with sucrose cushioning followed by ultracentrifugation

Abstract

Abstract Ginseng is a traditional medicine that has been used for millennia. Asian ginseng (Panax ginseng Meyer) has multiple pharmacological properties. To determine the efficacy of ginseng-derived exosome-like nanoparticles (GDEs), GDEs should be isolated and characterized. GDEs from P. ginseng were isolated via sequential centrifugation using 68 and 27% of sucrose cushioning followed by ultracentrifugation. The size distribution, zeta potential, and morphology of GDEs were evaluated using DLS, NTA, and TEM. GDEs were quantified by measuring the total RNA and protein concentration. The stability of the freeze-dried GDEs was evaluated for 90 days. Uniform and reproducible GDEs were successfully isolated with high yield and purity. GDEs were found to be spherical with an average diameter of 105.8 nm and a negative surface charge of − 20.7 mV. Substantial amounts of nanoparticles of GDEs were counted using NTA, with a concentration of 2.05 × 1013 particles/mL. According to the evaluation of their protein and total RNA concentration, the freeze-dried GDEs were stable for up to 60 days at room temperature. The results suggest the utility of a brief isolation protocol using sucrose double fractionation, which can be used for the mass production of plant-derived exosome-like nanoparticles. In addition, the stability of GDEs was maintained even after freeze-drying and storage for 60 days. This method has potential for application in the research for plant-derived exosome-like nanoparticles, where cost-effective exosome isolation with high yield and purity is an important step.

Keywords

• Ginseng
• Exosome
• Plant-derived exosome-like nanoparticle
• Ultracentrifugation
• Nanovesicles
• Isolation