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一个完全自动化的FAIMS-DIA基于质谱的蛋白质组管道

Luke Reilly¹, Erika Lara¹, Daniel Ramos¹

¹Center for Alzheimer's and Related Dementias (CARD), National Institute on Aging and National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA.

Cell reports methods

|September 20, 2023

概括

这项研究引入了一个精简的蛋白质组学管道,用于深度细胞蛋白质组覆盖. 使用FAIMS-DIA和Spectronaut的自动化工作流程,在iPSC衍生的神经元中识别了9000多种蛋白质,具有高精度和可重复性.

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:

深度蛋白质组覆盖对于理解细胞功能至关重要.
需要高通量和可扩展的蛋白质组学方法来进行全面的分析.
数据独立获取 (DIA) 为蛋白质组分析提供了优势.

研究的目的:

开发和验证一个标准化,高通量蛋白质组学管道.
优化DIA策略,以深入覆盖细胞蛋白质组.
评估FAIMS-DIA和Spectronaut用于蛋白质组分析的性能.

主要方法:

一个集成的,自动化的样本准备平台.
数据独立采集 (DIA) 与高场不对称波形离子移动谱学 (FAIMS) 相结合.
没有图书馆的DIA数据库搜索使用Spectronaut.

主要成果:

用FAIMS-DIA在-35V的单次补偿电压 (CV) 实现了最深的蛋白质覆盖.
"光谱飞行员"在直接DIA数据库搜索中表现出卓越的表现,量化了最多的蛋白质.
优化的FAIMS-DIA方法在iPSC衍生的神经元中识别了9000多种蛋白质,缺少值为10%以下,显示出高可重现性和准确性.

结论:

开发的"现成"蛋白质组学管道使深度和可扩展的细胞蛋白质组分析成为可能.
具有单个CV设置和Spectronaut的FAIMS-DIA是高通量蛋白质组的强大组合.
这种方法在复杂的生物样本上,与现有的DIA策略相比,提供了更高的性能.

关键词:

CP: 生物技术生物技术 CP:系统生物学系统生物学 DDA DDA DDA DDA DDA DDA DDA DDA DDA DDA DDA DDA DDA DDA DDA DDA DDA 时间时间时间时间他们很饥饿.自动化自动化自动化自动化搜索数据库搜索数据库搜索数据库搜索质谱测量质谱测量质谱测量质谱测量质量测量质谱测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量蛋白质组学蛋白质组学一次性射击MSMS

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