Automated Immunoprecipitation, Sample Preparation, and Individual Ion Mass Spectrometry Platform for Proteoforms.

Autor: Des Soye BJ; 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., McGee JP; 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.; ImmPro, Evanston, Illinois 60201, United States., Hollas MAR; 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., Forte E; Proteomics Center of Excellence, Northwestern University, Evanston, Illinois 60208, United States.; Comprehensive Transplant Center, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States., Fellers RT; 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., Melani RD; 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., Wilkins JT; 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.; Departments of Medicine (Cardiology) and Preventive Medicine (Epidemiology), Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, United States., Compton PD; Integrated Protein Technologies, San Jose, California 95134, United States., Kafader JO; 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., Kelleher NL; 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.
Jazyk: angličtina
Zdroj: Analytical chemistry [Anal Chem] 2024 Aug 14. Date of Electronic Publication: 2024 Aug 14.
DOI: 10.1021/acs.analchem.4c01962
Abstrakt: 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 (I 2 MS). 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 I 2 MS 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 I 2 MS spectra that contained a positive COVID-19 antibody response and 21 I 2 MS spectra that contained a negative COVID-19 antibody response. This high-throughput automated I 2 MS workflow can currently process over 100 samples per week and is general for making immunoprecipitation-MS workflows achieve proteoform resolution.
Databáze: MEDLINE
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