PMID: 9530562Apr 8, 1998Paper

Pathfinder selection: in situ isolation of novel antibodies

Immunotechnology : an International Journal of Immunological Engineering
J K OsbournK S Johnson

Abstract

To devise a novel method for targeted recovery of binding molecules from phage libraries. To assess the potential of the novel technique to the selection of human antibodies to specific cell surface antigens in situ, including carcinoembryonic antigen (CEA), E- and P-selectins, and to the selection of novel antibodies which recognize immobilized purified antigen. Recovery of these antibodies from a naive human scFv library was effected using a 'pathfinder' molecule. Monoclonal and polyclonal antibodies, as well as natural ligands can serve as pathfinders when conjugated directly or indirectly to horseradish peroxidase (HRP). In the presence of biotin tyramine these molecules catalyze biotinylation of phage binding in close proximity to the target antigen, allowing specific recovery of 'tagged' phage from the total population using streptavidin. In this way, phage binding to the target itself, or in its immediate proximity, are selectively recovered. This work demonstrates that an existing binding specificity can be used as a tool to select phage libraries in situ, obviating the need to purify or clone the target. The speed and technical simplicity of this method should find a wide range of applications to phage display librarie...Continue Reading

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Citations

Aug 1, 2000·Immunology Today·H R Hoogenboom, P Chames
Mar 21, 1998·Current Opinion in Biotechnology·A D Griffiths, A R Duncan
Apr 21, 1999·Current Opinion in Biotechnology·E V ShustaK D Wittrup
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Jun 13, 1998·Nature Biotechnology·T J VaughanP R Tempest
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Feb 28, 2001·Proceedings of the National Academy of Sciences of the United States of America·M A HuieJ D Marks
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Jan 10, 2014·Protein Engineering, Design & Selection : PEDS·Alexander MaaßHarald Kolmar
Jul 24, 2007·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Christa Mersich, Alois Jungbauer
Jul 21, 2004·Journal of Immunological Methods·Andrew R M Bradbury, James D Marks
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Apr 27, 2017·Protein Engineering, Design & Selection : PEDS·Séverine FagèteOliver Hartley
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Oct 30, 2020·Biochemistry·You-Chiun ChangKurt Yun Mou
May 10, 2008·Biochimie·Jean-Luc Jestin

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