Wang Chris, Wang Austin, Wei Fang, Wong David T W, Tu Michael
School of Dentistry, University of California, Los Angeles, CA, USA.
Methods Mol Biol. 2017;1660:367-376. doi: 10.1007/978-1-4939-7253-1_30.
In order to more fully elucidate the biogenesis of exosomes, a type of extracellular vesicles (EVs) and understand the role of EVs in disease processes, it is necessary to develop methods to capture EVs and induce the unloading of viable cargo. Traditionally, ultracentrifugation followed by chemical based EV lysis techniques is used to isolate these extracellular vesicles and release their internal content. Here, we describe a novel technique for capturing and releasing exosomal content through magnetic bead-based EV extraction coupled with electric field induced release and measurement (EFIRM). The usage of low-voltage electric fields allows the EV to be lysed without chemical treatment, and surface immobilized probes can allow for the rapid capture of the content of lysed EVs. EFIRM as an integrated EV lysing and analysis system offers great potential for the investigation of EVs in clinical and basic science contexts.
为了更全面地阐明外泌体（一种细胞外囊泡（EVs））的生物发生过程，并了解EVs在疾病进程中的作用，开发捕获EVs并诱导其有活力的货物卸载的方法很有必要。传统上，先进行超速离心，然后采用基于化学的EV裂解技术来分离这些细胞外囊泡并释放其内部成分。在此，我们描述了一种通过基于磁珠的EV提取结合电场诱导释放和测量（EFIRM）来捕获和释放外泌体内容物的新技术。低电压电场的使用使得EVs无需化学处理即可裂解，并且表面固定的探针能够快速捕获裂解后EVs的内容物。EFIRM作为一种集成的EV裂解和分析系统，在临床和基础科学领域对EVs的研究中具有巨大潜力。
