Absolute quantification of tumor antigens using embedded MHC-I isotopologue calibrants

Absolute quantification measurements (copies per cell) of peptide major histocompatibility complex (pMHC) antigens are necessary to inform targeted immunotherapy drug design; however, existing methods for absolute quantification have critical limitations. Here, we present a platform termed SureQuant...

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Detalles Bibliográficos
Autores principales: Stopfer, Lauren E., Gajadhar, Aaron S., Patel, Bhavin, Gallien, Sebastien, Frederick, Dennie T., Boland, Genevieve M., Sullivan, Ryan J., White, Forest M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2021
Materias:
Biological Sciences
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8449407/
https://www.ncbi.nlm.nih.gov/pubmed/34497125
http://dx.doi.org/10.1073/pnas.2111173118
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author Stopfer, Lauren E.
Gajadhar, Aaron S.
Patel, Bhavin
Gallien, Sebastien
Frederick, Dennie T.
Boland, Genevieve M.
Sullivan, Ryan J.
White, Forest M.
author_facet Stopfer, Lauren E.
Gajadhar, Aaron S.
Patel, Bhavin
Gallien, Sebastien
Frederick, Dennie T.
Boland, Genevieve M.
Sullivan, Ryan J.
White, Forest M.
author_sort Stopfer, Lauren E.
collection PubMed
description Absolute quantification measurements (copies per cell) of peptide major histocompatibility complex (pMHC) antigens are necessary to inform targeted immunotherapy drug design; however, existing methods for absolute quantification have critical limitations. Here, we present a platform termed SureQuant-IsoMHC, utilizing a series of pMHC isotopologues and internal standard-triggered targeted mass spectrometry to generate an embedded multipoint calibration curve to determine endogenous pMHC concentrations for a panel of 18 tumor antigens. We apply SureQuant-IsoMHC to measure changes in expression of our target panel in a melanoma cell line treated with a MEK inhibitor and translate this approach to estimate antigen concentrations in melanoma tumor biopsies.
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spelling pubmed-84494072021-10-04 Absolute quantification of tumor antigens using embedded MHC-I isotopologue calibrants Stopfer, Lauren E. Gajadhar, Aaron S. Patel, Bhavin Gallien, Sebastien Frederick, Dennie T. Boland, Genevieve M. Sullivan, Ryan J. White, Forest M. Proc Natl Acad Sci U S A Biological Sciences Absolute quantification measurements (copies per cell) of peptide major histocompatibility complex (pMHC) antigens are necessary to inform targeted immunotherapy drug design; however, existing methods for absolute quantification have critical limitations. Here, we present a platform termed SureQuant-IsoMHC, utilizing a series of pMHC isotopologues and internal standard-triggered targeted mass spectrometry to generate an embedded multipoint calibration curve to determine endogenous pMHC concentrations for a panel of 18 tumor antigens. We apply SureQuant-IsoMHC to measure changes in expression of our target panel in a melanoma cell line treated with a MEK inhibitor and translate this approach to estimate antigen concentrations in melanoma tumor biopsies. National Academy of Sciences 2021-09-14 2021-09-08 /pmc/articles/PMC8449407/ /pubmed/34497125 http://dx.doi.org/10.1073/pnas.2111173118 Text en Copyright © 2021 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Stopfer, Lauren E.
Gajadhar, Aaron S.
Patel, Bhavin
Gallien, Sebastien
Frederick, Dennie T.
Boland, Genevieve M.
Sullivan, Ryan J.
White, Forest M.
Absolute quantification of tumor antigens using embedded MHC-I isotopologue calibrants
title Absolute quantification of tumor antigens using embedded MHC-I isotopologue calibrants
title_full Absolute quantification of tumor antigens using embedded MHC-I isotopologue calibrants
title_fullStr Absolute quantification of tumor antigens using embedded MHC-I isotopologue calibrants
title_full_unstemmed Absolute quantification of tumor antigens using embedded MHC-I isotopologue calibrants
title_short Absolute quantification of tumor antigens using embedded MHC-I isotopologue calibrants
title_sort absolute quantification of tumor antigens using embedded mhc-i isotopologue calibrants
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8449407/
https://www.ncbi.nlm.nih.gov/pubmed/34497125
http://dx.doi.org/10.1073/pnas.2111173118
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