高通量蛋白质组和蛋白质组分析,用于甲固定式嵌入组织
Moe Haines1, John R Thorup1, Simone Gohsman1
1Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Molecular & cellular proteomics : MCP
|July 31, 2025
概括
本研究介绍了一种新的,自动化的工作流程,用于对甲固定,嵌 (FFPE) 组织的蛋白质组分析. 该方法有效地识别了数千种蛋白质和酸盐,在档案样本中推进了生物标志物发现.
科学领域:
- 生物化学 生物化学
- 蛋白质组学是指蛋白质组学.
- 分子生物学分子生物学
背景情况:
- 在将蛋白质组数据与临床结果联系起来时,固定在甲上,嵌入胺 (FFPE) 组织至关重要.
- 现有的FPE蛋白质组工作流程可能是耗时和劳动密集的.
研究的目的:
- 开发和优化FFPE组织蛋白质学的高吞吐量,部分自动化工作流.
- 为了实现深层次的蛋白质组覆盖和档案样本的定量稳定性.
- 用FFPE患者组织推进生物标志物发现.
主要方法:
- 以病理为导向的宏切割和使用自适应聚焦声学 (AFA) 超声波的无烯脱.
- 优化了S-Trap的消化和清理.
- 在Exploris 480,Orbitrap Astral和timsTOF HT平台上进行液体染色学-并联质谱学 (LC-MS/MS).
主要成果:
- 每个样本识别了8,000-10,000个独特的蛋白质,中位数CVs<20%.
- 在肺腺癌 (LUAD) 阻断FFPE方面表现出有效性,实现了深度蛋白质覆盖.
- 在LUAD FFPE组织中使用短梯度的Orbitrap Astral识别了多达10,000种蛋白质和11,000种酸盐.
结论:
- 开发的工作流是可扩展和高效的FPPE蛋白质组学.
- 它提供了深厚的蛋白质组覆盖范围和与现有方法相比的定量准确性.
- 这个平台促进了生物标志物发现和在档案组织样本中的蛋白质组研究.
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