Best practices for optimization and validation of flow cytometry‐based receptor occupancy assays. Issue 1 (1st December 2020)

• Record Type: Journal Article
• Authors: Hilt, Ed; Sun, Yongliang S.; McCloskey, Thomas W.; Eck, Steve; McIntosh, Thomas; Grugan, Katharine D.; Lanham, David F.; Standifer, Nathan; Green, Cherie; Litwin, Virginia; Stewart, Jennifer J.
• Is Part Of: Cytometry. Volume 100:Issue 1(2021)
• Page Start: 63
• Page End: 71
• Publication Date: 2020-12-01
• Subjects: drug development -- flow cytometry -- immunotherapeutics -- receptor occupancy -- validation; Flow cytometry -- Diagnostic use -- Periodicals; Cytodiagnosis -- Periodicals; 616.07582
• DOI: 10.1002/cyto.b.21970
• Languages: English
• ISSNs: 1552-4949

Abstract

Abstract: In the development of therapeutic compounds that bind cell surface molecules, it is critical to demonstrate the extent to which the drug engages its target. For cell‐associated targets, flow cytometry is well‐suited to monitor drug‐to‐target engagement through receptor occupancy assays (ROA). The technology allows for the identification of specific cell subsets within heterogeneous populations and the detection of nonabundant cellular antigens. There are numerous challenges in the design, development, and implementation of robust ROA. Among the most difficult challenges are situations where there is receptor modulation or when the target‐antigen is expressed at low levels. When the therapeutic molecules are bi‐specific and bind multiple targets, these challenges are increased. This manuscript discusses the challenges and proposes best practices for designing, optimizing, and validating ROA.