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Journal of Biomolecular Screening
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A Simple, No-Wash Cell Adhesion–Based High-Throughput Assay for the Discovery of Small-Molecule Regulators of the Integrin CD11b/CD18

Jun Y. Park

Nephrology Division, Leukocyte Biology and Inflammation Program, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA

M. Amin Arnaout

Nephrology Division, Leukocyte Biology and Inflammation Program, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA

Vineet Gupta

Nephrology Division, Leukocyte Biology and Inflammation Program, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, vineet_gupta{at}hms.harvard.edu

The leukocyte-specific integrin CD11b/CD18 plays a key role in the biological function of these cells and represents a validated therapeutic target for inflammatory diseases. Currently, the low affinity interaction between CD11b/CD18 integrin and its respective ligand poses a challenge in the development of cell-based adhesion assays for the high-throughput screening (HTS) environment. Here the authors describe a simple cell-based adhesion assay that can be readily used for HTS for the discovery of functional regulators of CD11b/CD18. The assay consistently produces acceptable Z' values (> 0.5) for HTS. After testing the assay using 2 established blocking antibodies as reference biologicals, the authors performed a proof-of-concept primary screen using a library of 6612 compounds and identified both agonist and antagonist hits. (Journal of Biomolecular Screening 2007:406-417)

Key Words: integrin • no-wash assay • cell adhesion assay • primary screen

Journal of Biomolecular Screening, Vol. 12, No. 3, 406-417 (2007)
DOI: 10.1177/1087057106299162
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