Abstract
Background
Camel milk is a source of exosomes with potential immunomodulatory and antioxidant benefits. However, the efficiency of exosome extraction from camel milk is crucial for maximizing their potential applications.
Objective
This study aims to optimize the exosome isolation process from camel milk to increase the yield and quality of the extracted exosomes.
Methods
We employed various pretreatment strategies prior to ultracentrifugation, including chymosin-assisted, isoelectric point (PI)-precipitation, and ethylenediaminetetraacetic acid (EDTA)-assisted methods. For the characterization of the isolated exosomes, we utilized nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and western blotting to evaluate size distribution, structural integrity, and specific exosomal protein markers, respectively.
Results
The chymosin-assisted technique yielded exosomes with a more intact and defined double-layered membrane structure, lower non-exosomal protein background, and a higher presence of the exosomal marker CD63 as evidenced by western blotting analysis (P < 0.05), compared with the other methodologies tested.
Conclusion
Chymosin pretreatment combined with ultracentrifugation significantly enhances the isolation of high-quality exosomes from camel milk, indicating that this approach may be the most effective for purifying exosomes for downstream biomedical applications. Further studies are encouraged to validate and refine this method for large-scale extraction.