title: A combinatorial approach to engineering a dual-specific metal switch antibody.
abstract: There is considerable interest in understanding how multiple binding events can be mediated through a single protein interface. Here, a synthetic library approach was developed to generate a novel dual-specific antibody. Using a combinatorial histidine-scanning phage display library, potential metal binding sites were introduced throughout an anti-RNase A antibody interface. Stepwise selection of RNase A and metal binding produced a dual-specific antibody that retained near wild-type affinity for its target antigen while acquiring a competitive metal binding site that is capable of controlling the antibody-antigen interaction. Structure analysis of the original antibody-RNase A complex suggested peripheral interface residues and loop flexibility are key contributors for obtaining the dual specificity.
authors.affiliation: Department of Chemistry and Biochemistry, Northern Illinois University, DeKalb, IL 60115, USA.
authors.forename: Sean W
authors.initials: SW
authors.lastname: Fanning
authors.forename: Megan L
authors.initials: ML
authors.lastname: Murtaugh
authors.forename: James R
authors.initials: JR
authors.lastname: Horn
mesh_terms: D018033:Antibodies, Bispecific / Q000737:chemistry* / Q000378:metabolism; D000915:Antibody Affinity; D000941:Antigens / Q000737:chemistry / Q000378:metabolism; D001666:Binding Sites, Antibody; D006639:Histidine / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008670:Metals / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D019151:Peptide Library; D015202:Protein Engineering; D012259:Ribonuclease, Pancreatic / Q000737:chemistry / Q000378:metabolism
chemical_list: D018033:Antibodies, Bispecific; D000941:Antigens; D008670:Metals; D019151:Peptide Library; D006639:Histidine; D012259:Ribonuclease, Pancreatic