Brown Peter N, Yin Hang
Drug Discovery Programme, Beatson Institute for Cancer Research, Bearsden, Glasgow, G61 1BD, UK.
Department of Chemistry & Biochemistry, University of Colorado Boulder, 3415 Colorado Ave, Boulder, CO, 80309, USA.
Methods Mol Biol. 2017;1660:91-103. doi: 10.1007/978-1-4939-7253-1_8.
Lipid particles, including exosomes (extracellular vesicles [EVs]), are released from cells in vivo and in vitro. The contents of these EVs can be indicative of disease and therefore be utilized in diagnostic biophysical or biochemical assays. To make use of EVs in this way, methods for purification and quantification are required which vary depending on their particles origin and concentration. This chapter provides an overview of EV purification techniques and provides detailed instructions on the purification of EVs by polymer based precipitation and subsequent quantification. The subsequent quantification and characterization of these EVs also presents a challenge, as limited methods are capable of detecting EVs due to their small size.
脂质颗粒,包括外泌体(细胞外囊泡[EVs]),可在体内和体外从细胞中释放出来。这些细胞外囊泡的内容物可指示疾病,因此可用于诊断性生物物理或生化检测。要以这种方式利用细胞外囊泡,就需要根据其颗粒来源和浓度的不同而采用不同的纯化和定量方法。本章概述了细胞外囊泡的纯化技术,并提供了基于聚合物沉淀法纯化细胞外囊泡及其后续定量的详细说明。这些细胞外囊泡的后续定量和表征也面临挑战,因为由于其尺寸小,能够检测到它们的方法有限。