Exosomes separation with aqueous two-phase systems from bovine milk

doi:10.1016/j.cjche.2024.12.016

中国化学工程学报 ›› 2025, Vol. 81 ›› Issue (5): 1-10.DOI: 10.1016/j.cjche.2024.12.016

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Exosomes separation with aqueous two-phase systems from bovine milk

Jingjing Sun, Ruoque Mao, Xiaoqian Fu, Shanjing Yao, Dongqiang Lin

Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang Key Laboratory of Smart Biomaterials, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310058, China

收稿日期:2024-10-15 修回日期:2024-11-28 接受日期:2024-12-30 出版日期:2025-05-28 发布日期:2025-03-08
通讯作者: Dongqiang Lin,E-mail:lindq@zju.edu.cn
基金资助:
This work was financially supported by the National Natural Science Foundation of China (22378350).

Exosomes separation with aqueous two-phase systems from bovine milk

Jingjing Sun, Ruoque Mao, Xiaoqian Fu, Shanjing Yao, Dongqiang Lin

Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang Key Laboratory of Smart Biomaterials, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310058, China

Received:2024-10-15 Revised:2024-11-28 Accepted:2024-12-30 Online:2025-05-28 Published:2025-03-08
Contact: Dongqiang Lin,E-mail:lindq@zju.edu.cn
Supported by:
This work was financially supported by the National Natural Science Foundation of China (22378350).

摘要/Abstract

摘要： The exosomes hold significant potential in disease diagnosis and therapeutic interventions. The objective of this study was to investigate the potential of aqueous two-phase systems (ATPSs) for the separation of bovine milk exosomes. The milk exosome partition behaviors and bovine milk separation were investigated, and the ATPSs and bovine milk whey addition was optimized. The optimal separation conditions were identified as 16% (mass) polyethylene glycol 4000, 10% (mass) dipotassium phosphate, and 1% (mass) enzymatic hydrolysis bovine milk whey. During the separation process, bovine milk exosomes were predominantly enriched in the interphase, while protein impurities were primarily found in the bottom phase. The process yielded bovine milk exosomes of 2.0 × 10¹¹ particles per ml whey with high purity (staining rate>90%, 7.01 × 10¹⁰ particles per mg protein) and high uniformity (polydispersity index <0.03). The isolated exosomes were characterized and identified by transmission electron microscopy, zeta potential and size distribution. The results demonstrated aqueous two-phase extraction possesses a robust capability for the enrichment and separation of exosomes directly from bovine milk whey, presenting a novel approach for the large-scale isolation of exosomes.

关键词: Aqueous two-phase extraction, Bovine milk exosomes, Separation, Purification

Abstract: The exosomes hold significant potential in disease diagnosis and therapeutic interventions. The objective of this study was to investigate the potential of aqueous two-phase systems (ATPSs) for the separation of bovine milk exosomes. The milk exosome partition behaviors and bovine milk separation were investigated, and the ATPSs and bovine milk whey addition was optimized. The optimal separation conditions were identified as 16% (mass) polyethylene glycol 4000, 10% (mass) dipotassium phosphate, and 1% (mass) enzymatic hydrolysis bovine milk whey. During the separation process, bovine milk exosomes were predominantly enriched in the interphase, while protein impurities were primarily found in the bottom phase. The process yielded bovine milk exosomes of 2.0 × 10¹¹ particles per ml whey with high purity (staining rate>90%, 7.01 × 10¹⁰ particles per mg protein) and high uniformity (polydispersity index <0.03). The isolated exosomes were characterized and identified by transmission electron microscopy, zeta potential and size distribution. The results demonstrated aqueous two-phase extraction possesses a robust capability for the enrichment and separation of exosomes directly from bovine milk whey, presenting a novel approach for the large-scale isolation of exosomes.

Key words: Aqueous two-phase extraction, Bovine milk exosomes, Separation, Purification

Jingjing Sun, Ruoque Mao, Xiaoqian Fu, Shanjing Yao, Dongqiang Lin. Exosomes separation with aqueous two-phase systems from bovine milk[J]. 中国化学工程学报, 2025, 81(5): 1-10.

Jingjing Sun, Ruoque Mao, Xiaoqian Fu, Shanjing Yao, Dongqiang Lin. Exosomes separation with aqueous two-phase systems from bovine milk[J]. Chinese Journal of Chemical Engineering, 2025, 81(5): 1-10.

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Exosomes separation with aqueous two-phase systems from bovine milk

Exosomes separation with aqueous two-phase systems from bovine milk

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