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Best practices for optimization and validation of flow cytometry‐based receptor occupancy assays
Author(s) -
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.
Publication year - 2021
Publication title -
cytometry part b: clinical cytometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 61
eISSN - 1552-4957
pISSN - 1552-4949
DOI - 10.1002/cyto.b.21970
Subject(s) - flow cytometry , computational biology , occupancy , computer science , drug development , identification (biology) , receptor , drug , biology , pharmacology , immunology , biochemistry , ecology , botany
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.