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Development and Evaluation of a FACS-Based Medium-Throughput Assay for HCV Entry Inhibitors

Pharmaceutical and Medicinal Chemistry, Saarland University, Saarbrücken, Germany

Bernd Kronenberger

Medizinische Klinik I, Frankfurt, Germany

Beatrice A.-M. Albrecht

Pharmaceutical and Medicinal Chemistry, Saarland University, Saarbrücken, Germany

Artur Kaul

Department of Molecular Virology, University of Heidelberg, Heidelberg, Germany

Anna-Lena Gamer

Department of Molecular Virology, University of Heidelberg, Heidelberg, Germany

Christian D. Klein

Pharmaceutical and Medicinal Chemistry, Saarland University, Saarbrücken, Germany, c.klein{at}uni-heidelberg.de

Rolf W. Hartmann

Pharmaceutical and Medicinal Chemistry, Saarland University, Saarbrücken, Germany, rwh{at}mx.uni-saarland.de

The interaction between the hepatitis C virus (HCV) envelope glycoprotein E2 and the human tetraspanin protein CD81 is one of the key events involved in HCV cell entry. Therefore, compounds that interfere with this interaction may be useful tools for basic research and potential drugs for the treatment of HCV infection. The authors describe a medium-throughput assay for ligands of the E2 binding site on the CD81 receptor. In the assay, human hepatoma cells are incubated with the test compounds and stained with a fluorescently labeled anti-CD81 antibody (JS81). Flow cytometry is used to detect the level of bound antibody, reflecting the inhibitory potencies of the compounds. Eighty percent of compounds active in the assay show efficacy in an infection assay using luciferase reporter genome in cell culture. Thus, the assay can be used as a fast screening system for inhibitors of interaction of viral E2 to host cell CD81-LELs. (Journal of Biomolecular Screening 2009;620-626)

Key Words: CD81-LEL • HCV entry inhibitors • HCV-E2 protein • protein-protein interaction inhibitors

This version was published on July 1, 2009

Journal of Biomolecular Screening, Vol. 14, No. 6, 620-626 (2009)
DOI: 10.1177/1087057109337161


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