Scholarlink

<p>Bovine milk-derived extracellular vesicles (BmEVs) have emerged as promising, biocompatible nanocarriers for drug delivery and therapeutic interventions. However, the lack of standardized and scalable isolation protocols remains a significant bottleneck for their industrial and clinical translation. In this study, we performed a multi-dimensional head-to-head comparison between traditional ultracentrifugation (UC) and tangential flow filtration (TFF) systems to evaluate their potential for large-scale BmEV manufacturing. We first evaluated the impact of TFF membrane pore sizes (300&#xa0;kDa and 750&#xa0;kDa) and flow rates on purification dynamics. The results suggest that TFF achieved isolation yields and quality comparable to UC while maintaining the essential physicochemical properties, including size, zeta potential, and morphology. Subsequent label-free data-independent acquisition (DIA) quantitative proteomics revealed that the TFF-750&#xa0;kDa approach closely replicated the global proteomic landscape of the gold-standard UC. Notably, the TFF-750&#xa0;kDa system showed enhanced preservation of transmembrane EV markers compared to UC, whereas the TFF-300&#xa0;kDa membrane led to substantial co-retention of milk protein impurities. Furthermore, in vitro biosafety assessments indicated that TFF-750&#xa0;kDa isolates exhibited efficient HepG2 cellular uptake, stability under simulated gastric fluid conditions, and favorable hemocompatibility with no significant cytotoxicity. These findings support the potential of the TFF-750&#xa0;kDa method as a promising and scalable alternative to UC for producing biocompatible and functional BmEV formulations.</p> Graphical abstract <p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Comparative profiling of tangential flow filtration and ultracentrifugation for the scalable production of bovine milk extracellular vesicles

  • 期刊论文

  • 发表日期：

    2026年5月13日

查看全文
  • Ling Zhang,
  • Ronglian Xu,
  • Fayin Wang,
  • Shihan Wang,
  • Xiaorong Fan,
  • Xinru Zhang,
  • Zhifang Zhou,
  • Zhimeng Wu,
  • Jie Shi

摘要

Bovine milk-derived extracellular vesicles (BmEVs) have emerged as promising, biocompatible nanocarriers for drug delivery and therapeutic interventions. However, the lack of standardized and scalable isolation protocols remains a significant bottleneck for their industrial and clinical translation. In this study, we performed a multi-dimensional head-to-head comparison between traditional ultracentrifugation (UC) and tangential flow filtration (TFF) systems to evaluate their potential for large-scale BmEV manufacturing. We first evaluated the impact of TFF membrane pore sizes (300 kDa and 750 kDa) and flow rates on purification dynamics. The results suggest that TFF achieved isolation yields and quality comparable to UC while maintaining the essential physicochemical properties, including size, zeta potential, and morphology. Subsequent label-free data-independent acquisition (DIA) quantitative proteomics revealed that the TFF-750 kDa approach closely replicated the global proteomic landscape of the gold-standard UC. Notably, the TFF-750 kDa system showed enhanced preservation of transmembrane EV markers compared to UC, whereas the TFF-300 kDa membrane led to substantial co-retention of milk protein impurities. Furthermore, in vitro biosafety assessments indicated that TFF-750 kDa isolates exhibited efficient HepG2 cellular uptake, stability under simulated gastric fluid conditions, and favorable hemocompatibility with no significant cytotoxicity. These findings support the potential of the TFF-750 kDa method as a promising and scalable alternative to UC for producing biocompatible and functional BmEV formulations.

Graphical abstract