Sancho-Albero María, Sebastián Víctor, Sesé Javier, Pazo-Cid Roberto, Mendoza Gracia, Arruebo Manuel, Martín-Duque Pilar, Santamaría Jesús
Department of Chemical Engineering, University of Zaragoza, 50018, Zaragoza, Spain.
Instituto de Nanociencia y Materiales de Aragón (INMA), CSIC-Universidad de Zaragoza, 50009, Zaragoza, Spain.
J Nanobiotechnology. 2020 Oct 22;18(1):150. doi: 10.1186/s12951-020-00701-7.
Exosomes are endocytic-extracellular vesicles with a diameter around 100 nm that play an essential role on the communication between cells. In fact, they have been proposed as candidates for the diagnosis and the monitoring of different pathologies (such as Parkinson, Alzheimer, diabetes, cardiac damage, infection diseases or cancer).
In this study, magnetic nanoparticles (FeONPs) were successfully functionalized with an exosome-binding antibody (anti-CD9) to mediate the magnetic capture in a microdevice. This was carried out under flow in a 1.6 mm (outer diameter) microchannel whose wall was in contact with a set of NdFeB permanent magnets, giving a high magnetic field across the channel diameter that allowed exosome separation with a high yield. To show the usefulness of the method, the direct capture of exosomes from whole blood of patients with pancreatic cancer (PC) was performed, as a proof of concept. The captured exosomes were then subjected to analysis of CA19-9, a protein often used to monitor PC patients.
Here, we describe a new microfluidic device and the procedure for the isolation of exosomes from whole blood, without any need of previous isolation steps, thereby facilitating translation to the clinic. The results show that, for the cases analyzed, the evaluation of CA19-9 in exosomes was highly sensitive, compared to serum samples.
外泌体是直径约100纳米的内吞细胞外囊泡,在细胞间通讯中起重要作用。事实上,它们已被提议作为诊断和监测不同疾病(如帕金森病、阿尔茨海默病、糖尿病、心脏损伤、传染病或癌症)的候选物。
在本研究中,磁性纳米颗粒(FeONPs)成功地用外泌体结合抗体(抗CD9)功能化,以介导在微器件中的磁性捕获。这是在一个外径为1.6毫米的微通道中流动条件下进行的,该微通道的壁与一组钕铁硼永磁体接触,在通道直径上产生一个强磁场,从而能够以高产率分离外泌体。为了证明该方法的实用性,作为概念验证,对胰腺癌(PC)患者的全血进行了外泌体的直接捕获。然后对捕获的外泌体进行CA19-9分析,CA19-9是一种常用于监测PC患者的蛋白质。
在此,我们描述了一种新的微流控装置以及从全血中分离外泌体的程序,无需任何先前的分离步骤,从而便于向临床转化。结果表明,对于所分析的病例,与血清样本相比,外泌体中CA19-9的评估具有高度敏感性。