Rapid isolation method for extracellular vesicles based on Fe3O4@ZrO2

Cuidie Ma; Zhihui Xu; Kun Hao; Lingling Fan; Wenqian Du; Zhan Gao; Chong Wang; Zheng Zhang; Ningxia Li; Qi Li; Qi Gao; Changyuan Yu

doi:10.3389/fbioe.2024.1399689

Rapid isolation method for extracellular vesicles based on Fe₃O₄@ZrO₂

Front Bioeng Biotechnol. 2024 Jul 9:12:1399689. doi: 10.3389/fbioe.2024.1399689. eCollection 2024.

Authors

Cuidie Ma^#¹, Zhihui Xu^#², Kun Hao², Lingling Fan², Wenqian Du², Zhan Gao³, Chong Wang⁴, Zheng Zhang⁵, Ningxia Li⁶, Qi Li⁷, Qi Gao², Changyuan Yu¹

Affiliations

¹ College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, China.
² Beijing Hotgen Biotech Co., Ltd., Beijing, China.
³ Department of Urology, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China.
⁴ Department of Clinical Laboratory, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
⁵ Department of Clinical Laboratory, The First Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
⁶ Department of Clinical Laboratory, The Second Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi, China.
⁷ Department of Clinical Laboratory, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China.

^# Contributed equally.

Abstract

Extracellular vesicles (EVs) are pivotal in intercellular communication, disease mechanisms. Despite numerous methods for EVs isolation, challenges persist in yield, purity, reproducibility, cost, time, and automation. We introduce a EVs isolation technique using Fe₃O₄@ZrO₂ beads, leveraging ZrO₂-phosphate interaction. The results indicated that EVs were efficiently separated from large volumes of samples in 30 minutes without preconcentration. Our method demonstrated capture efficiency (74%-78%) compared to ultracentrifugation, purity (97%), and reproducibility (0.3%-0.5%), with excellent linearity (R² > 0.99). EVs from urine samples showed altered expression of miRNAs. The logistic regression model achieved an AUC of 0.961, sensitivity of 0.92, and specificity of 0.94. With potential for automation, this magnetic bead-based method holds promise for clinical applications, offering an efficient and reliable tool for EVs research and clinical studies.

Keywords: ZrO2; extracellular vesicles; isolation; microRNAs; prostate cancer.

Grants and funding

The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This research was funded by the National Natural Science Foundation of China (No. 82174531), National High-Level Hospital Clinical Research Funding (XK2023-13), and the Scientific and Technological Research Project of the Xinjiang Production and Construction Corps (grant number 2022AB022).