Comparative analysis of exosome isolation methods using culture supernatant for optimum yield, purity and downstream applications
Comparative analysis of exosome isolation methods using culture supernatant for optimum yield, purity and downstream applications
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DOI：
10.1038/s41598-019-41800-2
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发表时间：
2019-03-29
影响因子：
4.6
通讯作者：
Singh, Ajay Pratap
中科院分区：
文献类型：
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作者：
Patel, Girijesh Kumar;Khan, Mohammad Aslam;Singh, Ajay Pratap
外泌体因其在病理生物学过程中的作用而受到极大关注，并被探索作为疾病诊断和管理的工具。因此，已经开发了基于不同原理的各种分离方法用于外泌体分离。在这里，我们比较了来自Invitrogen、101 Bio、和子和iZON沿着的四种试剂盒与传统的基于超离心的方法对于外泌体产率、纯度和质量的功效。使用细胞培养上清液作为外泌体的丰富来源，并测定外泌体数量、大小分布、zeta电位、标志物表达和RNA/蛋白质质量。Invitrogen试剂盒给出了最高的产率，但制剂显示出可能由于微泡共沉淀而导致的更宽的尺寸分布，并且具有最低的分散稳定性。其他准备工作表明，
Exosomes have received significant attention for their role in pathobiological processes and are being explored as a tool for disease diagnosis and management. Consequently, various isolation methods based on different principles have been developed for exosome isolation. Here we compared the efficacy of four kits from Invitrogen, 101Bio, Wako and iZON along with conventional ultracentrifugation-based method for exosome yield, purity and quality. Cell culture supernatant was used as an abundant source of exosomes, and exosome quantity, size-distribution, zeta-potential, marker-expression and RNA/protein quality were determined. The Invitrogen kit gave the highest yield but the preparation showed broader size-distribution likely due to microvesicle co-precipitation and had the least dispersion stability. Other preparations showed