微小的细胞外囊泡的染色学净化使用相亲系列对脂膜的相亲系列
Kanako Masaki1, Abo Bakr F Ahmed1,2, Takenori Ishida1
1Unit of Biotechnology, Graduate School of Integrated Sciences for Life, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-8530, Japan.
Biotechnology letters
|October 3, 2023
概括
一个EXOPUA亲和柱提供了小细胞外囊泡 (sEVs) 的高效,可扩展的净化. 与传统技术相比,这种染色学方法显著提高了sEV纯度和产量.
科学领域:
- 生物技术是生物技术.
- 细胞外囊泡研究研究
- 染色体学 染色体学 是一种染色学.
背景情况:
- 小细胞外囊泡 (sEVs) 是重要的生物标志物和治疗剂.
- 目前的净化方法,如微分超离心,耗时,可能会影响sEV的完整性.
- 需要快速,可扩展和可重复的sEV净化技术.
研究的目的:
- 评估ExoPUA亲和色谱列在净化小细胞外囊泡 (sEVs) 的有效性.
- 确定ExoPUA列是否提供了有效,快速,可扩展和可重复的sEV隔离方法.
主要方法:
- 使用了ExoUP A柱与快速性能液态色谱系统相结合.
- 采用绑定-化色谱协议,从培养上游生物中进行一阶段的sEV净化.
- 使用传输电子显微镜和标记分析 (CD9,CD63,TSG101,Calnexin,miR-21) 进行了净化的sEV的特征.
主要成果:
- 在单个步骤中实现了82 ± 16倍的sEV纯度增加,产量为38 ± 5%.
- 证实了正规sEV标记物 (CD9,CD63,TSG101) 和miRNA (miR-21) 的存在,没有Calnexin.
- 通过传输电子显微镜证明完整的sEV形态.
- 与差异超离心法相比,表现出更高的产量,比DEAE染色学更高的纯度.
结论:
- ExoPUA 列在绑定-出模式下有效地净化了 sEV.
- 在用氧化 (NaOH) 灭菌后,该柱是可重复使用的.
- 这种方法显示出可扩展和工业级别的sEV净化具有重大潜力.
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