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

Benjamin J. Des Soye; John P. McGee; Michael A.R. Hollas; Eleonora Forte; Ryan T. Fellers; Rafael D. Melani; John T. Wilkins; Philip D. Compton; Jared O. Kafader; Neil L. Kelleher

doi:10.1021/acs.analchem.4c01962

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

Benjamin J. Des Soye, John P. McGee, Michael A.R. Hollas, Eleonora Forte, Ryan T. Fellers, Rafael D. Melani, John T. Wilkins, Philip D. Compton, Jared O. Kafader, Neil L. Kelleher^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

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 (I²MS). 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 I²MS 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 I²MS spectra that contained a positive COVID-19 antibody response and 21 I²MS spectra that contained a negative COVID-19 antibody response. This high-throughput automated I²MS workflow can currently process over 100 samples per week and is general for making immunoprecipitation-MS workflows achieve proteoform resolution.

Original language	English (US)
Journal	Analytical Chemistry
DOIs	https://doi.org/10.1021/acs.analchem.4c01962
State	Accepted/In press - 2024

Funding

This study was funded by the National Institute of Health under a grant from the National Institute of General Medical Sciences P41 GM108569 (N.L.K.); Walder Foundation grant number SCI16; the NIH Office of Director award S10 OD025194 (P.D.C.); the Northwestern Medicine Dr. Michael M. Abecassis Transplant Innovation Endowment Grant; NCI CCSG P30 CA060553 (awarded to the Robert H. Lurie Comprehensive Cancer Center, N.L.K., E.F.). The authors would like to acknowledge the Northwestern University Clinical and Translational Sciences Institute and the Northwestern Memorial Foundation for their financial support of the Serology Study research effort.

ASJC Scopus subject areas

Analytical Chemistry

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title = "Automated Immunoprecipitation, Sample Preparation, and Individual Ion Mass Spectrometry Platform for Proteoforms",

abstract = "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 (I2MS). 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 I2MS 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 I2MS spectra that contained a positive COVID-19 antibody response and 21 I2MS spectra that contained a negative COVID-19 antibody response. This high-throughput automated I2MS workflow can currently process over 100 samples per week and is general for making immunoprecipitation-MS workflows achieve proteoform resolution.",

author = "{Des Soye}, {Benjamin J.} and McGee, {John P.} and Hollas, {Michael A.R.} and Eleonora Forte and Fellers, {Ryan T.} and Melani, {Rafael D.} and Wilkins, {John T.} and Compton, {Philip D.} and Kafader, {Jared O.} and Kelleher, {Neil L.}",

note = "Publisher Copyright: {\textcopyright} 2024 American Chemical Society.",

year = "2024",

doi = "10.1021/acs.analchem.4c01962",

language = "English (US)",

journal = "Analytical Chemistry",

issn = "0003-2700",

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T1 - Automated Immunoprecipitation, Sample Preparation, and Individual Ion Mass Spectrometry Platform for Proteoforms

AU - Des Soye, Benjamin J.

AU - McGee, John P.

AU - Hollas, Michael A.R.

AU - Forte, Eleonora

AU - Fellers, Ryan T.

AU - Melani, Rafael D.

AU - Wilkins, John T.

AU - Compton, Philip D.

AU - Kafader, Jared O.

AU - Kelleher, Neil L.

PY - 2024

Y1 - 2024

N2 - 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 (I2MS). 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 I2MS 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 I2MS spectra that contained a positive COVID-19 antibody response and 21 I2MS spectra that contained a negative COVID-19 antibody response. This high-throughput automated I2MS workflow can currently process over 100 samples per week and is general for making immunoprecipitation-MS workflows achieve proteoform resolution.

AB - 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 (I2MS). 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 I2MS 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 I2MS spectra that contained a positive COVID-19 antibody response and 21 I2MS spectra that contained a negative COVID-19 antibody response. This high-throughput automated I2MS workflow can currently process over 100 samples per week and is general for making immunoprecipitation-MS workflows achieve proteoform resolution.

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

Abstract

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ASJC Scopus subject areas

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