CLIN CHEM LAB MED:六种适用于临床实验室的血清外泌体分离方法对细胞因子分析的影响的比较

2019-11-17 MedSci MedSci原创

外泌体是可以在血液中检测到由细胞释放的纳米囊泡。外泌体含有多种分子,如细胞因子,因此可作为疾病的生物标志物。本工作的目的是比较六种不同的适用于临床实验室的商业试剂盒的效率和纯度,以及它们在后续细胞因子分析中的作用。<span lang="EN-US" style="font-size:12.0pt;mso-bidi-font-size:14.0pt;font-family:&quot;Calibr

外泌体是可以在血液中检测到由细胞释放的纳米囊泡。外泌体含有多种分子,如细胞因子,因此可作为疾病的生物标志物。本工作的目的是比较六种不同的适用于临床实验室的商业试剂盒的效率和纯度,以及它们在后续细胞因子分析中的作用。

本研究的血清外泌体取自10名志愿者,随后使用6种商业试剂盒:exoEasyExoQuickExo-spinME kitExoQuick PlusExo-Flow。通过纳米颗粒跟踪分析定量分析外泌体的浓度和大小分布。Western blot检测外泌体标志物CD63CD9TSG101。用比浊法测定ApoB和白蛋白。采用Luminex测定S100A9CXCL5CXCL12

研究结果表明,不同试剂盒之间得到的颗粒浓度变化范围为100倍左右。 ExoQuick and Exo-Flow除外,不同试剂盒间提取的颗粒浓度无相关性。ExoQuick PlusexoEasy获得的exosome纯度最高,而MEExoQuick最低。除 Exo-Flow and ME提取的外泌体提取物外,分析的所有外泌体提取物中均含有白蛋白,所有外泌体提取物均含有ApoB。细胞因子检测因使用的纯化试剂盒不同而不同,不同试剂盒获得的样品之间细胞因子浓度无相关性。

研究表明,样品和商业试剂盒的类型都会影响外泌体分离的效率和纯度。此外,外泌体纯化方法对细胞因子的检测和定量能力有重要影响。

原始出处:

Mónica MacíasVera RebmannComparison of six commercial serum exosome isolation methods suitable for clinical laboratories. Effect in cytokine analysis

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