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Wellcome Open Res. 2017 Sep 7;2:82. doi: 10.12688/wellcomeopenres.12058.1. eCollection 2017.

A new panel of epitope mapped monoclonal antibodies recognising the prototypical tetraspanin CD81.

Author information

Institute of Immunity and Transplantation, Division of Infection and Immunity, , University College London, London, NW3 2PF, UK.
Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, B15 2TT, UK.
Institute for Microbiology and Infection, School of Biosciences, University of Birmingham, Birmingham, B15 2TT, UK.
Centre for Biomedical Resear, Burnet Institute, Melbourne, VIC, 3004, Australia.
School of Life and Health Sciences, Aston University, Birmingham, B4 7ET, UK.
Nuffield Department of Medicine, University of Oxford, Oxford, OX3 7BN, UK.
Contributed equally


Background: Tetraspanins are small transmembrane proteins, found in all higher eukaryotes, that compartmentalize cellular membranes through interactions with partner proteins. CD81 is a prototypical tetraspanin and contributes to numerous physiological and pathological processes, including acting as a critical entry receptor for hepatitis C virus (HCV). Antibody engagement of tetraspanins can induce a variety of effects, including actin cytoskeletal rearrangements, activation of MAPK-ERK signaling and cell migration. However, the epitope specificity of most anti-tetraspanin antibodies is not known, limiting mechanistic interpretation of these studies. Methods: We generated a panel of monoclonal antibodies (mAbs) specific for CD81 second extracellular domain (EC2) and performed detailed epitope mapping with a panel of CD81 mutants. All mAbs were screened for their ability to inhibit HCV infection and E2-CD81 association. Nanoscale distribution of cell surface CD81 was investigated by scanning electron microscopy. Results: The antibodies were classified in two epitope groups targeting opposing sides of EC2. We observed a wide range of anti-HCV potencies that were independent of their epitope grouping, but associated with their relative affinity for cell-surface expressed CD81. Scanning electron microscopy identified at least two populations of CD81; monodisperse and higher-order assemblies, consistent with tetraspanin-enriched microdomains. Conclusions: These novel antibodies provide well-characterised tools to investigate CD81 function, including HCV entry, and have the potential to provide insights into tetraspanin biology in general.


CD81; hepatitis C virus; tetraspanin

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