The isolation of specific subpopulations of extracellular vesicles (EVs) based on their expression of surface markers poses a significant challenge due to their nanoscale size (< 800 nm), their heterogeneous surface marker expression, and the vast number of background EVs present in clinical specimens (10¹⁰–10¹² EVs/mL in blood). Highly parallelized nanomagnetic sorting using track etched magnetic nanopore (TENPO) chips has achieved precise immunospecific sorting with high throughput and resilience to clogging. However, there has not yet been a systematic study of the design parameters that control the trade-offs in throughput, target EV recovery, and ability to discard background EVs in this approach. We combine finite-element simulation and experimental characterization of TENPO chips to elucidate design rules to isolate EV subpopulations from blood. We demonstrate the utility of this approach by reducing device background > 10× relative to prior published designs without sacrificing recovery of the target EVs by selecting pore diameter, number of membranes placed in series, and flow rate. We compare TENPO-isolated EVs to those of gold-standard methods of EV isolation and demonstrate its utility for wide application and modularity by targeting subpopulations of EVs from multiple models of disease including lung cancer, pancreatic cancer, and liver cancer.

由于细胞外囊泡 (EV) 的纳米级尺寸 (< 800 nm)、异质表面标记表达以及临床样本中存在大量背景 EV，因此根据表面标记的表达来分离特定的细胞外囊泡 (EV) 亚群提出了重大挑战（血液中 10 ¹⁰ –10 ¹²  EVs/mL）。使用径迹蚀刻磁性纳米孔 (TENPO) 芯片的高度并行纳米磁性分选实现了精确的免疫特异性分选，具有高通量和抗堵塞能力。然而，目前还没有对控制这种方法中吞吐量、目标 EV 恢复和丢弃背景 EV 的能力权衡的设计参数进行系统研究。我们结合 TENPO 芯片的有限元模拟和实验表征来阐明从血液中分离 EV 亚群的设计规则。我们通过选择孔径、串联放置的膜数量和流速，通过相对于先前发布的设计减少> 10倍的设备背景来证明这种方法的实用性，而不牺牲目标电动汽车的回收率。我们将 TENPO 分离的 EV 与 EV 分离的金标准方法进行比较，并通过针对多种疾病模型（包括肺癌、胰腺癌和肝癌）的 EV 亚群来证明其广泛应用和模块化的实用性。