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Directed selection of MIP-1α neutralizing CCR5 antibodies from a phage display human antibody library

Nature Biotechnology volume 16, pages 778–781 (1998)Cite this article

Abstract

The seven trans-membrane chemokine receptor CCR-5 is a coreceptor for macrophage tropic HIV-1 strains. CCR-5 responds to a number of chemokines, including macrophage inflammatory protein (MIP)-1α. We describe the use of MIP-1α in a biotin tyramine-mediated proximity selection to guide the selection of CCR-5-specific phage antibodies from a large phage display human library. Proximity based selections resulted in a population of antibodies recognizing CCR-5 on primary CD4+ lymphocytes, none of which blocked MIP-1α binding to cells. The selected population of phage antibodies were subsequently used as guide molecules for a second phase of selection that was carried out in the absence of MIP-1α. This generated a panel of CCR-5-binding antibodies, of which around 20% inhibited MIP-1α binding to CD4+. The single chain Fvs (scFv) generated by this step-back selection procedure also inhibited MIP-1α-mediated calcium signaling.

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Authors and Affiliations

  1. Cambridge Antibody Technology Limited, The Science Park, Melbourn, Cambridgeshire, SC8 6JJ, U.K.

    Jane K. Osbourn, John C. Earnshaw, Kevin S. Johnson & John McCafferty

  2. IRIBHN and Service de Genetique Medicale, University Libre De Bruxelles, Campus Erasme, Bruxelles, Belgium

    Mark Parmentier

  3. Molecular Devices, Sunnyvale, CA, 94089

    Véronique Timmermans

Authors
  1. Jane K. Osbourn
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  2. John C. Earnshaw
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  3. Kevin S. Johnson
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  4. Mark Parmentier
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  5. Véronique Timmermans
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  6. John McCafferty
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Correspondence to Jane K. Osbourn.

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Osbourn, J., Earnshaw, J., Johnson, K. et al. Directed selection of MIP-1α neutralizing CCR5 antibodies from a phage display human antibody library. Nat Biotechnol 16, 778–781 (1998). https://doi.org/10.1038/nbt0898-778

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