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Selection of human antibody fragments by phage display

Nature Protocols volume 2, pages 3001–3008 (2007)Cite this article

Abstract

Here, we describe a protocol for the selection of human antibody fragments using repertoires displayed on filamentous bacteriophage. Antigen-specific clones are enriched by binding to immobilized antigen, followed by elution and repropagation of phage. After multiple rounds of binding selection, specific clones are identified by ELISA. This article provides an overview of phage display and antibody technology, as well as detailed protocols for the immobilization of antigen, the selection of repertoires on purified or complex antigens and the identification of binders.

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Figure 1: Phage display of antibody fragments.
Figure 2: Selection of antibody repertoire.
Figure 3: Monoclonal phage ELISA.
Figure 4: Anticipated results for monoclonal phage ELISA.

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Acknowledgements

Protocols are based on methods originally developed by D. Christ and others in Greg Winter's group at the MRC Laboratory of Molecular Biology and were modified in our laboratory at the Garvan Institute. D.C. was a Fellow of Trinity College (Cambridge University) from 2002 to 2006. This work was funded by a Cancer Institute NSW grant (07/RFG/1-12) to D.C. and a Cancer Institute NSW Early Career Fellowship to F.S. We thank G. Winter for helpful discussions.

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  1. Garvan Institute of Medical Research, 384 Victoria Road, Darlinghurst, 2010, Sydney New South Wales, Australia

    Carol M Y Lee, Niccolo Iorno, Frederic Sierro & Daniel Christ

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  1. Carol M Y Lee
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Correspondence to Daniel Christ.

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Lee, C., Iorno, N., Sierro, F. et al. Selection of human antibody fragments by phage display. Nat Protoc 2, 3001–3008 (2007). https://doi.org/10.1038/nprot.2007.448

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