Enrichment of bovine milk-derived extracellular vesicles using surface-functionalized cellulose nanofibers
Jonne Ukkola, Feby Wijaya Pratiwi, Santeri Kankaanpää, Seyedamirhosein Abdorahimzadeh, Mohammad Karzarjeddi, Prateek Singh, Artem Zhyvolozhnyi, Olha Makieieva, Sirja Viitala, Anatoly Samoylenko, Hely Häggman, Seppo Vainio, Çağlar Elbüken, Henrikki Liimatainen
Abstract
The isolation of extracellular vesicles (EVs) from milk, a complex mixture of colloidal structures having a comparable size to EVs, is challenging. Although ultracentrifugation (UC) has been widely used for EV isolation, this has significant limitations, including a long processing time at high g-force conditions and large sample volume requirements. We introduced a new approach based on nature nanoentities cellulose nanofibers (CNFs) and short time and low g-force centrifugation to isolate EVs from various milk fractions. The flexible and entangled network of CNFs forms nanoporous, which entraps the EVs. Further, positively charged CNFs interact with anionic EVs through an electrostatic attraction, promoting their isolation with efficiency comparable with UC. The functionality and toxicity of isolated milk EVs were tested in Caco2 cells. Overall, the newly developed approach provides straightforward isolation and biocompatibility and preserves the natural properties of the isolated EVs, enabling further applications.