Des Soye Benjamin J, McGee John P, Hollas Michael A R, Forte Eleonora, Fellers Ryan T, Melani Rafael D, Wilkins John T, Compton Philip D, Kafader Jared O, Kelleher Neil L
Departments of Chemistry and Molecular Biosciences, the Chemistry of Life Processes Institute, Northwestern University, Evanston, Illinois 60208, United States.
Proteomics Center of Excellence, Northwestern University, Evanston, Illinois 60208, United States.
Anal Chem. 2024 Aug 14. doi: 10.1021/acs.analchem.4c01962.
Charge detection mass spectrometry (CDMS) is a well-established technique that provides direct mass spectral outputs regardless of analyte heterogeneity or molecular weight. Over the past few years, it has been demonstrated that CDMS can be multiplexed on Orbitrap analyzers utilizing an integrated approach termed individual ion mass spectrometry (IMS). To further increase adaptability, robustness, and throughput of this technique, here, we present a method that utilizes numerous integrated equipment components including a Kingfisher system, SampleStream platform, and Q Exactive mass spectrometer to provide a fully automated workflow for immunoprecipitation, sample preparation, injection, and subsequent IMS acquisition. This automated workflow has been applied to a cohort of 58 test subjects to determine individualized patient antibody responses to SARS-CoV-2 antigens. Results from a range of serum donors include 37 subject IMS spectra that contained a positive COVID-19 antibody response and 21 IMS spectra that contained a negative COVID-19 antibody response. This high-throughput automated IMS workflow can currently process over 100 samples per week and is general for making immunoprecipitation-MS workflows achieve proteoform resolution.
电荷检测质谱（CDMS）是一种成熟的技术，无论分析物的异质性或分子量如何，都能提供直接的质谱输出。在过去几年中，已经证明CDMS可以在轨道阱分析仪上进行多路复用，采用一种称为单离子质谱（IMS）的集成方法。为了进一步提高该技术的适应性、稳健性和通量，在此，我们提出一种方法，该方法利用包括翠鸟系统、SampleStream平台和Q Exactive质谱仪在内的众多集成设备组件，为免疫沉淀、样品制备、进样以及随后的IMS采集提供一个全自动工作流程。这种自动化工作流程已应用于58名受试对象的队列，以确定个体患者对SARS-CoV-2抗原的抗体反应。一系列血清供体的结果包括37个含有阳性COVID-19抗体反应的受试对象IMS谱图和21个含有阴性COVID-19抗体反应的IMS谱图。这种高通量自动化IMS工作流程目前每周可处理超过100个样品，并且对于使免疫沉淀-质谱工作流程实现蛋白异构体分辨率具有通用性。
