title: Antibody library selection by the {beta}-lactamase protein fragment complementation assay.
abstract: Protein fragment complementation assay (PCA) is based on the interaction between two protein partners (e.g. target antigen and antibody), which are genetically fused to the two halves of a dissected marker protein. Binding of the two partners reassembles the marker protein and hence reconstitutes its activity. In this work we have developed the first application of beta-lactamase-based PCA for the isolation of single chain Fv fragments (scFvs) binding to the human receptor RON from a naïve library. Specific scFvs with the ability to immunoprecipitate could be isolated after a single round of PCA selection from an scFv repertoire previously pre-selected by phage display. Furthermore, the PCA was used to successfully map the epitopes recognized by the selected scFvs by screening them against a small library of random RON fragments.
authors.affiliation: Department of Medical Sciences, Interdisciplinary Research Centre on Autoimmune Diseases, University of Eastern Piedmont, Novara.
authors.forename: Paola
authors.initials: P
authors.lastname: Secco
authors.forename: Elena
authors.initials: E
authors.lastname: D'Agostini
authors.forename: Roberto
authors.initials: R
authors.lastname: Marzari
authors.forename: Marta
authors.initials: M
authors.lastname: Licciulli
authors.forename: Roberto
authors.initials: R
authors.lastname: Di Niro
authors.forename: Sara
authors.initials: S
authors.lastname: D'Angelo
authors.forename: Andrew R M
authors.initials: AR
authors.lastname: Bradbury
authors.forename: Umberto
authors.initials: U
authors.lastname: Dianzani
authors.forename: Claudio
authors.initials: C
authors.lastname: Santoro
authors.forename: Daniele
authors.initials: D
authors.lastname: Sblattero
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000915:Antibody Affinity / Q000235:genetics; D000941:Antigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D018604:Epitope Mapping; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D051379:Mice; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D019151:Peptide Library; D040081:Protein Array Analysis / Q000379:methods*; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000235:genetics* / Q000378:metabolism
chemical_list: D000906:Antibodies; D000941:Antigens; D010446:Peptide Fragments; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; C080835:RON protein; D020794:Receptor Protein-Tyrosine Kinases; D001618:beta-Lactamases