Direct isolation and characterization of circulating exosomes from biological samples using magnetic nanowires

Journal of Nanobiotechnology

卷:17

Mihye Choi¹  Jiyun Lim¹  Youngnam Cho¹  HyungJae Lee¹  Eun Sook Lee²  Young-Ho Kim³  Ji-Youn Han⁴ 

[1] Biomarker Branch, National Cancer Center;

[2] Division of Breast Cancer, National Cancer Center;

[3] Division of Clinical Research, Rare Cancer Branch, National Cancer Center;

[4] Division of Lung Cancer, National Cancer Center;

关键词: Plasma;    Exosome;    Magnetic nanowire;    Lung cancer;    Breast cancer;   

DOI  :  10.1186/s12951-018-0433-3

来源: DOAJ

【 摘 要 】

Abstract Background 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. Methods 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. Results 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. Conclusions 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.

【 授权许可】

Unknown