A multiplexed assay for quantifying immunomodulatory proteins supports correlative studies in immunotherapy clinical trials

Author:

Whiteaker, Jeffrey R

Zhao, Lei

Schoenherr, Regine M

Huang, Dongqing

Lundeen, Rachel A

Voytovich, Ulianna

Kennedy, Jacob J

Ivey, Richard G

Lin, Chenwei

Murillo, Oscar D

Lorentzen, Travis D

Colantonio,Simona

Caceres,Tessa

Roberts,Rhonda

Knotts,Joseph

Reading,Joshua

Perry,Candice

Richardson, Christopher W

Garcia-Buntley,Stephanie

Bocik,William

Hewitt, Stephen M

Chowdhury, Shrabanti

Vandermeer, Jackie

Smith, Stephen D

Gopal, Ajay K

Ramchurren, Nirasha

Fling, Steven P

Wang, Pei

Paulovich, Amanda G
Author Address

Clinical Research Division, Fred Hutchinson Cancer Center, Seattle, WA, United States., Cancer Research Technology Program, Antibody Characterization Lab, Frederick National Laboratory for Cancer Research, Frederick, MD, United States., Experimental Pathology Laboratory, Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institute of Health, Bethesda, MD, United States., Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, United States., Division of Medical Oncology, Department of Internal Medicine, University of Washington, Seattle, WA, United States., Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center, Seattle, WA, United States.,

Abstract:

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. 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. 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. Copyright © 2023 Whiteaker, Zhao, Schoenherr, Huang, Lundeen, Voytovich, Kennedy, Ivey, Lin, Murillo, Lorentzen, Colantonio, Caceres, Roberts, Knotts, Reading, Perry, Richardson, Garcia-Buntley, Bocik, Hewitt, Chowdhury, Vandermeer, Smith, Gopal, Ramchurren, Fling, Wang and Paulovich.

See More

Author Address