An input-controlled model system for identification of MHC bound peptides enabling laboratory comparisons of immunopeptidome experiments.

ABSTRACT: Characterization of MHC-bound peptides by mass spectrometry (MS) is an essential technique for immunologic studies. Many efforts have been made to quantify the number of MHC-presented ligands by MS and to define the limits of detection of a specific MHC ligand. However, these experiments are often complex and comparisons across different laboratories are challenging. Therefore, we compared and orthogonally validated quantitation of peptide:MHC complexes by radioimmunoassay and flow cytometry using TCR mimic antibodies in three model systems to establish a method to control the experimental input of peptide MHC:complexes for MS analysis. Following isolation of MHC-bound peptides we identified and quantified an MHC ligand of interest with high correlation to the initial input. We found that

SUBMITTER: Klatt MG 

PROVIDER: S-EPMC7484941 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature