Targeted Mass Spectrometry Enables Multiplexed Quantification of Immunomodulatory Proteins in Clinical Biospecimens

Front Immunol. 2021 Nov 11:12:765898. doi: 10.3389/fimmu.2021.765898. eCollection 2021.

Abstract

Immunotherapies are revolutionizing cancer care, producing durable responses and potentially cures in a subset of patients. However, response rates are low for most tumors, grade 3/4 toxicities are not uncommon, and our current understanding of tumor immunobiology is incomplete. While hundreds of immunomodulatory proteins in the tumor microenvironment shape the anti-tumor response, few of them can be reliably quantified. To address this need, we developed a multiplex panel of targeted proteomic assays targeting 52 peptides representing 46 proteins using peptide immunoaffinity enrichment coupled to multiple reaction monitoring-mass spectrometry. We validated the assays in tissue and plasma matrices, where performance figures of merit showed over 3 orders of dynamic range and median inter-day CVs of 5.2% (tissue) and 21% (plasma). A feasibility study in clinical biospecimens showed detection of 48/52 peptides in frozen tissue and 38/52 peptides in plasma. The assays are publicly available as a resource for the research community.

Keywords: cancer; correlative biomarkers; immuno-MRM; immunotherapy; mass spectrometry.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antibodies / analysis
  • Antibodies / immunology
  • Blotting, Western
  • Cell Line, Tumor
  • Chromatography, Liquid / methods*
  • HeLa Cells
  • Humans
  • Jurkat Cells
  • MCF-7 Cells
  • Mass Spectrometry / methods*
  • Peptides / analysis*
  • Peptides / blood
  • Peptides / immunology
  • Proteome / analysis*
  • Proteome / genetics
  • Proteome / immunology
  • Proteomics / methods*
  • RNA-Seq / methods
  • Reproducibility of Results
  • Specimen Handling / methods*

Substances

  • Antibodies
  • Peptides
  • Proteome