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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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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DOI: https://doi.org/10.1038/nbt0898-778
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