从不同的生物流体中分离出来的细胞外囊泡的度评估方法的比较
Gleb O Skryabin1, Adel D Enikeev1, Anastasiia A Beliaeva1
1Blokhin National Medical Research Center of Oncology, Moscow, 115522, Russia.
Biochemistry. Biokhimiia
|November 2, 2025
概括
准确的细胞外囊泡 (EV) 量化具有挑战性. 这项研究对各种生物流体的三种方法进行了比较,揭示了依赖于方法的差异,并强调了在EV研究中需要采用多因素方法.
科学领域:
- 生物医学研究生物医学研究
- 纳米技术 纳米技术
- 分析化学 分析化学
背景情况:
- 准确量化细胞外囊泡 (EVs) 对生物医学研究至关重要,但仍然具有挑战性.
- 由于样本中的非膀纳米粒子和生物污染物,现有的方法往往不可靠.
- 从子宫吸入物和胃汁等来源的EVs的定量评估在很大程度上是未经探索的.
研究的目的:
- 为了比较分析三种EV量化方法:总蛋白质含量,纳米粒子跟踪分析 (NTA) 和酶活性.
- 评估这些方法在从各种生物流体中分离的EV中,包括血,液,子宫吸附物,胃汁和癌细胞调节介质.
- 在EV量化中识别方法特定的局限性和取决于样本的变化.
主要方法:
- 从血,液,子宫吸附剂,胃汁和条件介质 (卵巢和非小细胞肺癌细胞) 中分离EVs.
- 使用总蛋白质含量测量进行量化.
- 使用纳米粒子追踪分析 (NTA) 进行量化.
- 使用酶活性试验 (商业FluoroCet套件) 进行量化.
主要成果:
- 这三种方法都显示出来自条件介质的EVs的强相关性,验证了它们用于用净化样本进行体外研究的使用.
- 在血,液和子宫吸附剂的方法之间观察到差异,可能是由于非膀纳米颗粒.
- 来自胃汁的EV显示了蛋白质含量和雌激酶活性之间的强烈相关性,表明其独特的组成.
结论:
- 选择EV量化方法显著影响结果,并取决于生物流体来源.
- 为了可靠的EV量化,需要考虑样本来源和方法限制的多因素方法.
- 这些发现为开发标准化的EV量化协议提供了基础,特别是对于临床样本.
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