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Digital Microfluidics for Automated Proteomic Processing
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自动免疫沉,样品制备和个人离子质谱平台为蛋白质形式
Benjamin J Des Soye1,2, John P McGee1,2,3, Michael A R Hollas1,2
1Departments of Chemistry and Molecular Biosciences, the Chemistry of Life Processes Institute, Northwestern University, Evanston, Illinois 60208, United States.
Analytical chemistry
|August 15, 2024
概括
电荷检测质谱 (CDMS) 已自动化,用于高通量分析. 这种单个离子质谱 (I2MS) 工作流准确地确定了58名受试者的SARS-CoV-2抗体反应.
科学领域:
- 质谱测量质量谱测量
- 免疫学 免疫学 免疫学
- 蛋白质组学是指蛋白质组学.
背景情况:
- 电荷检测质谱 (CDMS) 提供直接的质谱输出,无论分析物的特征如何.
- 个人离子质谱 (I2MS) 代表了 Orbitrap 分析仪上的多重化 CDMS 的进步.
研究的目的:
- 开发一个完全自动化的 immunoprecipitation 和随后的 I2MS 获取工作流.
- 提高I2MS分析的适应性,稳定性和吞吐量.
主要方法:
- 集成金鱼系统,SampleStream平台和Q Exactive质谱仪,用于自动化样品准备和I2MS.
- 应用自动化工作流来分析58个人的队列中对SARS-CoV-2抗原的抗体反应.
主要成果:
- 成功确定了个性化的患者抗体对SARS-CoV-2抗原的反应.
- 在37名受试者中确定了COVID-19抗体的阳性反应,在21名受试者中确定了阴性反应.
- 一个高通量工作流的演示,能够每周处理100多个样本.
结论:
- 开发的自动化I2MS工作流显著提高了免疫沉-MS的吞吐量和稳定性.
- 这种方法可以在免疫沉-MS工作流程中实现蛋白形状分辨率.
- 该工作流是有效的评估患者的抗体反应,如SARS-CoV-2抗原分析所示.
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