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Making antibody fragments using phage display libraries

Abstract

To by-pass hybridoma technology and animal immunization, we are trying to build antibodies in bacteria by mimicking features of immune selection1. Recently we used fd phage2 to display antibody fragments fused to a minor coat protein3,4, allowing enrichment of phage with antigen3. Using a random combinatorial library of the rearranged heavy (VH) and kappa (Vk) light chains5– from mice immune to the hapten 2-phenyloxazol-5-one (phOx), we have now displayed diverse libraries of antibody fragments on the surface of fd phage. After a single pass over a hapten affinity column, fd phage with a range of phOx binding activities were detected, at least one with high affinity (dissociation constant, Kd = 10-8 M). A second pass enriched for the strong binders at the expense of the weak. The binders were encoded by V genes similar to those found in anti-phOx hybridomas but in promiscuous combinations (where the same V gene is found with several different partners). By combining a promiscuous VH or VK gene with diverse repertoires of partners to create hierarchical libraries, we elicited many more pairings with strong binding activities. Phage display offers new ways of making antibodies from V-gene libraries, altering V-domain pairings and selecting for antibodies with good affinities.

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Clackson, T., Hoogenboom, H., Griffiths, A. et al. Making antibody fragments using phage display libraries. Nature 352, 624–628 (1991). https://doi.org/10.1038/352624a0

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