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一个样品制备程序为基于异标签的单细胞蛋白质学.

Consuelo Marín-Vicente1,2, Enrique Calvo3,4, José Manuel Rodríguez3

  • 1Cardiovascular Proteomics Group, Spanish National Centre for Cardiovascular Research (CNIC), Madrid, Spain. cmarin@cnic.es.

Methods in molecular biology (Clifton, N.J.)
|June 21, 2024
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概括
此摘要是机器生成的。

本研究提出了一种使用质谱学的单细胞蛋白质组学方法. 该协议提高了分析有限的生物样本的灵敏度,包括难以分离的细胞.

关键词:
质谱测量质量谱测量多重样本准备策略多重样本准备策略轨道探测器 Eclipse 质谱仪质谱仪单细胞蛋白质组学 单细胞蛋白质组学坦德姆群体标签 坦德姆群体标签三混合体质谱仪三混合体质谱仪

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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:

  • 传统的基于质谱的蛋白质组学需要大量的输入材料,限制了其应用.
  • 单细胞蛋白质组学需要超高的灵敏度,因为微小的样本数量.
  • 现有的方法在分析罕见或难以分离的细胞类型时面临挑战.

研究的目的:

  • 建立一个一般的工作流程,为单细胞质谱学为基础的蛋白质组学准备细胞.
  • 用载体蛋白质组适应等离体标签策略,以克服灵敏度的限制.
  • 为手工分离的细胞,特别是像心肌细胞这样的大细胞提供适用的协议.

主要方法:

  • 使用基于等标签的多重复合策略.
  • 结合载体蛋白质来增强信号检测.
  • 开发用于质谱分析的一般细胞准备工作流程.
  • 适应手动细胞隔离技术的协议.

主要成果:

  • 展示了基于单细胞质谱的蛋白质组学的一个强大的协议.
  • 该方法有效地解决了单细胞分析固有的灵敏度限制.
  • 工作流可以适应具有挑战性的细胞类型,包括那些不能接受标准排序的细胞类型.

结论:

  • 提出的协议显著提高了单细胞蛋白质组学的可行性.
  • 这种方法扩大了质谱仪在细胞生物学研究中的应用范围.
  • 该方法为分析珍贵或罕见细胞种群提供了有价值的工具.