Hong Chang-Sook, Funk Sonja, Whiteside Theresa L
University of Pittsburgh School of Medicine, University of Pittsburgh Cancer Institute, 5117 Centre Avenue, Suite 1.27, Pittsburgh, PA, 15213, USA.
Methods Mol Biol. 2017;1633:257-265. doi: 10.1007/978-1-4939-7142-8_16.
A method for exosome isolation from human plasma was developed for rapid, high-throughput processing of plasma specimens obtained from patients with cancer. This method removes the bulk of plasma proteins associated with exosomes and can be used for comparative examinations of exosomes and their content in serial specimens of patients' plasma, allowing for monitoring changes in exosome numbers, profiles, and functions in the course of cancer progression or during therapy. The plasma-derived exosomes can be recovered in quantities sufficient for the characterization of their morphology by transmission electron microscopy (TEM), size and concentration by qNano, protein/lipid ratios, nucleic acid extraction, molecular profiling by Western blots or immune arrays, and functional assays.
开发了一种从人血浆中分离外泌体的方法,用于对癌症患者的血浆样本进行快速、高通量处理。该方法去除了与外泌体相关的大部分血浆蛋白,可用于对患者血浆系列样本中的外泌体及其内容物进行比较检查,从而监测癌症进展过程或治疗期间外泌体数量、特征和功能的变化。从血浆中获得的外泌体数量足以通过透射电子显微镜(TEM)表征其形态,通过qNano测定其大小和浓度,测定蛋白质/脂质比率,进行核酸提取,通过蛋白质印迹或免疫阵列进行分子分析以及进行功能测定。