Extending the Comprehensiveness of Immunopeptidome Analyses Using Isobaric Peptide Labeling.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
Read More Add to Saved list
  • Additional Information
    • Source:
      Publisher: American Chemical Society Country of Publication: United States NLM ID: 0370536 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-6882 (Electronic) Linking ISSN: 00032700 NLM ISO Abbreviation: Anal Chem Subsets: MEDLINE
    • Publication Information:
      Original Publication: Washington, American Chemical Society.
    • Subject Terms:
      Histocompatibility Antigens Class I/*metabolism ; Molecular Probes/*chemistry ; Peptides/*analysis ; Tandem Mass Spectrometry/*methods; Animals ; Antibodies/immunology ; Cell Line, Tumor ; Chromatography, High Pressure Liquid ; Histocompatibility Antigens Class I/immunology ; Humans ; Immunoprecipitation ; Ion Mobility Spectrometry ; Mice ; Mice, Inbred NOD ; Peptides/chemistry ; Succinimides/chemistry ; Transplantation, Heterologous
    • Abstract:
      Defining the repertoire of peptides presented by the major histocompatibility complex class I (MHC I) is a key step toward the identification of relevant antigens for cancer immunotherapy. However, the identification of cancer-specific antigens is a significant analytical challenge in view of their low abundance and low mutational load found in most primary cancer specimens. Here, we describe the application of isobaric peptide labeling with tandem mass tag (TMT) to improve the detection of the MHC I peptides. Isobaric peptide labeling was found to promote the formation of multiply charged ions and to enhance the formation of b-type fragment ions, thus resulting in a 50% improvement of MHC I peptide identification. The gain in sensitivity obtained using TMT labeling enabled the detection of low-abundance MHC I peptides including tumor-specific antigens (TSAs) and minor histocompatibility antigens (MiHAs). We further demonstrate the application of this approach to quantify MiHAs presented by B-cell lymphocytes and determined their expression levels by LC-MS/MS using both synchronous precursor selection (SPS) and high-field asymmetric waveform ion mobility spectrometry (FAIMS).
    • Accession Number:
      0 (Antibodies)
      0 (Histocompatibility Antigens Class I)
      0 (Molecular Probes)
      0 (Peptides)
      0 (Succinimides)
      MJE3791M4T (N-hydroxysuccinimide)
    • Publication Date:
      Date Created: 20200606 Date Completed: 20210211 Latest Revision: 20210211
    • Publication Date:
      20221213
    • Accession Number:
      10.1021/acs.analchem.0c01545
    • Accession Number:
      32502341