期刊
NANOSCALE
卷 10, 期 29, 页码 14280-14289出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c8nr02871k
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资金
- National Natural Science Foundation of China (NSFC) [21571134, 31771037, 81471775]
- National Key Research and Development Program of China [2016YFC1102701]
- Project in the Science and Technology Support Program of Sichuan Province [18ZDYF0823]
- Project of Huimin Technology Research and Development of Chengdu [2015-HM01-00379-SF]
The study of exosomes has great significance for cancer diagnosis and prognosis, in which damage-free isolation of exosomes is a critically important step due to their extremely low concentration in body fluids. Herein, immunoaffinitive superparamagnetic nanoparticles (IS-NPs) were successfully constructed by combining antibodies with superparamagnetic nanoparticles through host-guest interactions between beta-cyclodextrin (beta-CD) and 4-aminoazobenzene (AAB). Exosomes in cell-culture supernatants or body fluids can be efficiently captured by IS-NPs and then mildly eluted by competitive host molecules, alpha-CD. Approximately 80% of the exosomes were captured by the IS-NPs from the model samples containing varied concentrations of exosomes (10(11)-10(7) exosomes per mL). After elution, the release efficiency of the captured exosomes was as high as 86.5%, as calculated from the extracted RNA content. A comparison between the exosomes isolated by our IS-NPs, conventional ultracentrifugation (UC), polyethylene glycol (PEG)-based precipitation, and a commercial kit was conducted to examine the performance of the IS-NP exosome isolation. The particle-to-protein ratio of IS-NP captured exosomes (8.8 +/- 1.3 x 10(9)) was eight-fold higher than that of the UC (1.1 +/- 0.4 x 10(9)) and more than two-fold that of the PEG-based precipitation (3.7 +/- 0.8 x 10(9)) and commercial kit (3.2 +/- 0.8 x 10(9)) isolated ones, revealing that IS-NPs yielded more exosomes with higher purity than all the other three approaches. Cellular uptake experiments demonstrated that the exosomes captured by IS-NPs have retained their structural and functional integrities, and their broad applications in biomedical areas could therefore be expected. Promisingly, this host-guest interaction involving immunoaffinity magnetic particle system could offer a new way for fast, high-efficiency and high-purity exosome isolation and elution, further benefitting exosomes' applications in desired areas.
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