Lee Hyo Joo, Lee Jinkwan, Kim Namheon, Shin Yong
Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Republic of Korea.
Infusiontech, 38 Heungan-daero, 427 Beon-gil, Dongan-gu, Anyang-si 14059, Republic of Korea.
Int J Mol Sci. 2025 Jul 16;26(14):6843. doi: 10.3390/ijms26146843.
Conventional methods for isolating extracellular vesicles (EVs) are often limited by long processing times, a low purity, and a reliance on specialized equipment. To overcome these challenges, we developed the DeSEI (amine-functionalized Diatomaceous earth-based Syringe platform for EV Isolation), a novel platform employing low-cost, amine-functionalized diatomaceous earth (ADe) within a simple syringe-filter system. The capture mechanism leverages the electrostatic interaction between the positively charged ADe and the negatively charged EV surface, enabling a rapid and efficient isolation. The optimized 30 min protocol yields intact EVs with morphology, size, and protein markers comparable to those from ultracentrifugation, ensuring minimal cellular contamination. Notably, DeSEI exhibited a nearly 60-fold higher recovery efficiency of EV-derived miRNA compared to ultracentrifugation. The platform further proved its versatility with a rapid one-step miRNA extraction protocol and a user-friendly cartridge format. The direct miRNA extraction capability is particularly advantageous for a streamlined biomarker analysis, while the cartridge design illustrates a clear pathway toward developing point-of-care diagnostic tools. The DeSEI offers a promising alternative to existing methods for EV-based research by providing a combination of speed, simplicity, and procedural flexibility that does not require specialized equipment.
传统的细胞外囊泡（EVs）分离方法通常受到处理时间长、纯度低以及依赖专业设备的限制。为了克服这些挑战，我们开发了DeSEI（用于EV分离的胺功能化硅藻土基注射器平台），这是一种在简单的注射器过滤系统中采用低成本胺功能化硅藻土（ADe）的新型平台。捕获机制利用带正电荷的ADe与带负电荷的EV表面之间的静电相互作用，实现快速高效的分离。优化后的30分钟方案可产生完整的EVs，其形态、大小和蛋白质标志物与超速离心法得到的相当，确保细胞污染最小。值得注意的是，与超速离心法相比，DeSEI对EV衍生的miRNA的回收效率高出近60倍。该平台通过快速的一步式miRNA提取方案和用户友好的试剂盒形式进一步证明了其通用性。直接提取miRNA的能力对于简化生物标志物分析特别有利，而试剂盒设计为开发即时诊断工具指明了一条清晰的途径。DeSEI通过提供速度、简单性和程序灵活性的组合，且无需专业设备，为基于EV的研究提供了一种有前景的替代现有方法的方案。
