title: Generating Conformation and Complex-Specific Synthetic Antibodies.
abstract: Phage display is commonly used to identify and isolate binders from large combinatorial libraries. Here we present phage selection protocols enabling generation of synthetic antibodies capable of recognizing multiprotein complexes and conformational states. The procedure describes stages of the experiment design, optimization, and screening, as well as provides the framework for building downstream assays with an end goal of isolating bioactive antibodies for future therapeutic use. The methods described are also applicable to screening directly on cells and can be ported to other in vitro directed evolution systems utilizing non-immunoglobulin scaffolds.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, KCBD Rm.3240G, 900 East 57th Street, Chicago, IL, 60637, USA. mpaduch@recombinant-antibodies.org.
authors.forename: Marcin
authors.initials: M
authors.lastname: Paduch
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, KCBD Rm.3240G, 900 East 57th Street, Chicago, IL, 60637, USA.|Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL, 60637, USA.
authors.forename: Anthony A
authors.initials: AA
authors.lastname: Kossiakoff
mesh_terms: D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D000918:Antibody Specificity; D006801:Humans; D008968:Molecular Conformation; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000276:immunology; D019151:Peptide Library
chemical_list: D000906:Antibodies; D046912:Multiprotein Complexes; D019151:Peptide Library
keywords: Antibody phage display; Conformation specific; Synthetic antibody