Protocol

Human Monoclonal Antibodies

Volume 1060 of the series Methods in Molecular Biology pp 277-295

Phage Display Technology for Human Monoclonal Antibodies

  • Cecilia DeantonioAffiliated withDepartment of Health Sciences and Interdisciplinary Research Center for Autoimmune Diseases (IRCAD), University of Eastern Piedmont
  • , Diego CotellaAffiliated withDepartment of Health Sciences and Interdisciplinary Research Center for Autoimmune Diseases (IRCAD), University of Eastern Piedmont
  • , Paolo MacorAffiliated withDepartment of Life Sciences, University of Trieste
  • , Claudio SantoroAffiliated withDepartment of Health Sciences and Interdisciplinary Research Center for Autoimmune Diseases (IRCAD), University of Eastern Piedmont
  • , Daniele SblatteroAffiliated withDepartment of Health Sciences and Interdisciplinary Research Center for Autoimmune Diseases (IRCAD), University of Eastern Piedmont

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Abstract

During the last 15 years in vitro technologies opened powerful routes to combine the generation of large libraries together with fast selection procedures to identify lead candidates. One of the commonest methods is based on the use filamentous phages. Antibodies (Abs) can be displayed successfully on the surface of phage by fusing the coding sequence of the antibody variable (V) regions to the phage minor coat protein pIII. By creating large libraries, antibodies with affinities comparable to those obtained using traditional hybridomas technology can be selected by a series of cycles of selection on antigen. As in this system antibody genes are cloned simultaneously with selection they can be easily further engineered for example by increasing their affinity (to levels unobtainable in the immune system), modulating their specificity or their effector function (by recloning into a full-length immunoglobulin scaffold). This chapter describes the basic protocols for antibody library construction, handling, and selection.

Key words

Phage display Antigens Monoclonal antibody High-throughput scFv