Open AccessAbsolute quantification of tumor antigens using embedded MHC-I isotopologue calibrants
Author(s) -
Lauren Stopfer,
Aaron S. Gajadhar,
Bhavin Patel,
Sébastien Gallien,
Dennie T. Frederick,
Genevieve M. Boland,
Ryan J. Sullivan,
Forest M. White
Publication year - 2021
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.2111173118
Subject(s) - isotopologue , major histocompatibility complex , antigen , mass spectrometry , computational biology , immunotherapy , chemistry , melanoma , cancer research , biology , immunology , chromatography , immune system , molecule , organic chemistry
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.