Biomarker Branch, National Cancer Center, 323 Ilsan-ro, Ilsan-dong-gu, Goyang, Gyeonggi, 10408, South Korea.
Department of Cancer Biomedical Science, Graduate School of Cancer Science and Policy, 323 Ilsan-ro, Ilsan-dong-gu, Goyang, Gyeonggi, 10408, South Korea.
J Nanobiotechnology. 2019 Jan 7;17(1):1. doi: 10.1186/s12951-018-0433-3.
Tumor-derived exosomes are gaining attention as important factors that facilitate communication between neighboring cells and manipulate cellular processes associated with cancer development or progression. The conventional techniques for the isolation and detection of exosomes face several limitations, restricting their clinical applications. Hence, a highly efficient technique for the isolation and identification of exosomes from biological samples may provide critical information about exosomes as biomarkers and improve our understanding of their unique role in cancer research. Here, we describe the use of antibody cocktail-conjugated magnetic nanowires to isolate exosomes from plasma of breast and lung cancer patients.
The isolated exosomes were characterized based on size and concentration using nanoparticle tracking analysis. Levels of exosomal proteins were measured by bicinchoninic acid assay and enzyme-linked immunosorbent assay. Morphology was visualized by transmission electron microscopy. Immunoblotting (Western blotting) was used to detect the presence of exosomal markers.
The use of antibody cocktail-conjugated magnetic nanowires resulted in approximately threefold greater yield when compared to the conventional methods. The elongated feature of nanowires significantly improved the efficiency of exosome isolation, suggesting its potential to be translated in diverse clinical applications, including cancer diagnosis and treatment.
The nanowire-based method allows rapid isolation of homogeneous population of exosomes with relatively high yield and purity from even small amounts of sample. These results suggest that this method has the potential for clinical applications requiring highly purified exosomes for the analysis of protein, lipid, mRNA, and miRNA.
肿瘤来源的外泌体作为促进相邻细胞间通讯的重要因素而受到关注，并能调控与癌症发生或进展相关的细胞过程。传统的外泌体分离和检测技术存在一些局限性，限制了其临床应用。因此，一种高效的从生物样本中分离和鉴定外泌体的技术，可能会为外泌体作为生物标志物提供关键信息，并增进我们对其在癌症研究中独特作用的理解。在这里，我们描述了使用抗体偶联磁纳米线从乳腺癌和肺癌患者的血浆中分离外泌体的方法。
使用纳米颗粒跟踪分析基于大小和浓度对分离的外泌体进行表征。通过双缩脲法和酶联免疫吸附测定法测量外泌体蛋白的水平。通过透射电子显微镜观察形态。免疫印迹（Western blot）用于检测外泌体标志物的存在。
与传统方法相比，抗体偶联磁纳米线的使用使产量增加了约三倍。纳米线的长形特征显著提高了外泌体的分离效率，表明其在包括癌症诊断和治疗在内的各种临床应用中有转化的潜力。
该基于纳米线的方法允许从即使少量的样本中快速分离出具有相对高产量和纯度的同质外泌体群体。这些结果表明，该方法有可能用于需要高度纯化的外泌体进行蛋白质、脂质、mRNA 和 miRNA 分析的临床应用。
