Table_1_A multiplexed assay for quantifying immunomodulatory proteins supports correlative studies in immunotherapy clinical trials.xlsx (66.25 kB)

Table_1_A multiplexed assay for quantifying immunomodulatory proteins supports correlative studies in immunotherapy clinical trials.xlsx

dataset

posted on 2023-05-02, 04:23 authored by Jeffrey R. Whiteaker, Lei Zhao, Regine M. Schoenherr, Dongqing Huang, Rachel A. Lundeen, Ulianna Voytovich, Jacob J. Kennedy, Richard G. Ivey, Chenwei Lin, Oscar D. Murillo, Travis D. Lorentzen, Simona Colantonio, Tessa W. Caceres, Rhonda R. Roberts, Joseph G. Knotts, Joshua J. Reading, Candice D. Perry, Christopher W. Richardson, Sandra S. Garcia-Buntley, William Bocik, Stephen M. Hewitt, Shrabanti Chowdhury, Jackie Vandermeer, Stephen D. Smith, Ajay K. Gopal, Nirasha Ramchurren, Steven P. Fling, Pei Wang, Amanda G. Paulovich

Introduction

Immunotherapy is an effective treatment for a subset of cancer patients, and expanding the benefits of immunotherapy to all cancer patients will require predictive biomarkers of response and immune-related adverse events (irAEs). To support correlative studies in immunotherapy clinical trials, we are developing highly validated assays for quantifying immunomodulatory proteins in human biospecimens.

Methods

Here, we developed a panel of novel monoclonal antibodies and incorporated them into a novel, multiplexed, immuno-multiple reaction monitoring mass spectrometry (MRM-MS)-based proteomic assay targeting 49 proteotypic peptides representing 43 immunomodulatory proteins.

Results and discussion

The multiplex assay was validated in human tissue and plasma matrices, where the linearity of quantification was >3 orders of magnitude with median interday CVs of 8.7% (tissue) and 10.1% (plasma). Proof-of-principle demonstration of the assay was conducted in plasma samples collected in clinical trials from lymphoma patients receiving an immune checkpoint inhibitor. We provide the assays and novel monoclonal antibodies as a publicly available resource for the biomedical community.