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Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

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Published on: November 17, 2019

对于无标签单细胞蛋白质组的简单和可访问的工作流程

Ximena Sanchez-Avila¹, Thy Truong¹, Xiaofeng Xie¹

¹Department of Chemistry and Biochemistry, Brigham Young University, Provo, Utah 84602, United States.

Journal of the American Society for Mass Spectrometry

|August 18, 2023

概括

一个新的,低成本的HP D100单细胞分剂器可以实现可访问的单细胞蛋白质组学 (SCP). 这个商业平台提供了精确的细胞分发和快速样本准备,扩大了SCP对研究人员的可访问性.

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Last Updated: May 23, 2026

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科学领域:

生物化学生化学
分析化学分析化学
分子生物学分子生物学

背景情况:

单细胞蛋白质组学 (SCP) 提供了独特的生物学见解,但往往受到高成本和专用设备的限制.
目前用于SCP的商业平台昂贵,需要大量的技术专业知识,阻碍了广泛采用.

研究的目的:

评估HP D100单细胞分剂器作为一种低成本,可访问的工具,用于无标签的单细胞蛋白质组学 (SCP).
评估使用HP D100用于SCP样本准备的精度,可重复性和蛋白质覆盖率.

主要方法:

使用HP D100单细胞分剂器,将单细胞分离和准备在384个井的PCR板中,以获得无标签的SCP.
采用细胞和试剂分配后的离心分离步骤,以优化单细胞回收和样品完整性.
使用家庭包装和商业纳米LC列分析单细胞样本.

主要成果:

惠普D100在200nL至2μL的剂量中表现出高精度和可重复性.
经过优化离心后,大约97%的分配井产生了与单细胞一致的蛋白质覆盖.
使用HP D100和井板的完全商业化工作流程提供了快速,易于使用的SCP样品准备,与纳米井方法相比,无损失蛋白质覆盖.

结论:

惠普D100单细胞分剂器,结合商业井板和一步处理,为SCP提供了快速,用户友好和成本效益的工作流.
这种方法显著提高了单细胞蛋白质组学对更广泛的实验室的可访问性.
虽然商业列减少了蛋白质识别,但整体商业平台为SCP样本准备和分离提供了可行的替代方案.