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

ABSTRACT: SureQuant-MHC utilizes a series of isotopologues and internal standard-triggered targeted mass spectrometry to generate an embedded multipoint calibration curve to determine the endogenous pMHC concentrations for a panel of 18 antigen targets. We apply this methiod to profile changes in pMHC expression levels in melanoma cell lines treated with binimetinib, a MEK inhibitr, and traslate the approach to profiling antigen levels in human tumor punch biopsies.

INSTRUMENT(S): Orbitrap Exploris 480, Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Melanocyte, Skin

SUBMITTER: Lauren Stopfer

LAB HEAD: Forest White

PROVIDER: PXD024917 | Pride | 2021-09-21

REPOSITORIES: Pride

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

Stopfer Lauren E LE Gajadhar Aaron S AS Patel Bhavin B Gallien Sebastien S Frederick Dennie T DT Boland Genevieve M GM Sullivan Ryan J RJ White Forest M FM

Proceedings of the National Academy of Sciences of the United States of America 20210901 37

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 concentra ...[more]

PMID: 34497125

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