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使用nPOPOP进行多重化单细胞蛋白质组的大量并行样本制备.

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科学领域:

  • 蛋白质组学是指蛋白质组学.
  • 质谱测量质量谱测量
  • 细胞生物学 细胞生物学

背景情况:

  • 使用质谱法 (MS) 的单细胞蛋白质组学提供了高的特异性和灵敏性.
  • 目前的方法在分析大量单个细胞时面临吞吐量限制.

研究的目的:

  • 开发一种高通量方法,用于单细胞蛋白质组样本的制备.
  • 为了使成千上万个单细胞的并行处理用于多重性多发性硬化症分析.

主要方法:

  • 在玻璃幻灯片上使用纳米升体积液滴开发了纳米蛋白质样品制备 (nPOP).
  • 实施nPOP与plexDIA (非同位体标签) 和同位体标签用于多重MS.
  • 利用一个细胞分配机器人 (CellenONE) 和随时可用的耗材.

主要成果:

  • 使用plexDIA. 精确量化每个人细胞的3,000-3,700个蛋白质.
  • 分析了1,827个单细胞,每天>1,000个细胞,800-1,200个蛋白质/细胞深度,使用异标签.
  • nPOP协议需要1-2天的时间来准备>3,000个单细胞.

结论:

  • nPOP显著增加了单细胞蛋白质组的吞吐量.
  • 该协议灵活,可扩展,适用于各种MS方法.
  • 质量控制软件 (QuantQC R包) 支持强大的数据分析和扩展.