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

Jisu Kim,Jianxin Wang,Yeon-Hee Lee,Il Keun Kwon,Youn Kyung Kim

doi:10.1007/s42452-022-04943-y

Jisu Kim, Jianxin Wang + Show 3 more

Open Access

https://doi.org/10.1007/s42452-022-04943-y

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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.

Highlights

Asian ginseng (Panax ginseng Meyer) is known as the king of herbs
We evaluated whether a novel sucrose cushioning method and sequential centrifugation could be used to obtain homogeneous and reproducible ginseng-derived exosome-like nanoparticles (GDEs) with high yield and purity
After centrifugation at a high velocity, the ginseng extracellular vesicles (EVs) were purified between the sucrose cushion layers (Fig. 2c)

Summary

Introduction

Ginseng has multiple pharmacological properties in cardiovascular diseases, diabetes, and cancer, and has been used to promote immune function, central nervous system function, relieve stress, and enhance bone remodeling [1]. It has been used in traditional medicine for the treatment of diseases in East. Asian countries for millennia, and until recently, it has become one of the most effective herbs 2. Numerous molecular targets for ginseng have been identified in recent years [5, 6], it is still not clear which factors are responsible for the therapeutic effects of ginseng

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