title: High throughput functional epitope mapping: revisiting phage display platform to scan target antigen surface.
abstract: Antibody engineering must be accompanied by mapping strategies focused on identifying the epitope recognized by each antibody to define its unique functional identity. High throughput fine specificity determination remains technically challenging. We review recent experiences aimed at revisiting the oldest and most extended display technology to develop a robust epitope mapping platform, based on the ability to manipulate target-derived molecules (ranging from the whole native antigen to antigen domains and smaller fragments) on filamentous phages. Single, multiple and combinatorial mutagenesis allowed comprehensive scanning of phage-displayed antigen surface that resulted in the identification of clusters of residues contributing to epitope formation. Functional pictures of the epitope(s) were thus delineated in the natural context. Successful mapping of antibodies against interleukin-2, epidermal growth factor and its receptor, and vascular endothelial growth factor showed the versatility of these procedures, which combine the accuracy of site-directed mutagenesis with the high throughput potential of phage display.
authors.affiliation: a Systems Biology Department ; Center of Molecular Immunology ; La Habana , Cuba.
authors.forename: Gertrudis
authors.initials: G
authors.lastname: Rojas
authors.forename: Yaima
authors.initials: Y
authors.lastname: Tundidor
authors.forename: Yanelys Cabrera
authors.initials: YC
authors.lastname: Infante
mesh_terms: D000941:Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D061505:Cell Surface Display Techniques / Q000379:methods*; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D006801:Humans; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed / Q000379:methods; D019151:Peptide Library; D017434:Protein Structure, Tertiary
chemical_list: D000941:Antigens; D000939:Epitopes; D019151:Peptide Library
keywords: Abs, antibodies; Ag, antigen; EGF; EGF receptor; EGF, epidermal growth factor; EGFR, EGF receptor; ELISA, enzyme-linked immunosorbent assay; IL-2; IL-2, interleukin-2; PCR, polymerase chain reaction; VEGF; VEGF, vascular endothelial growth factor; aa, amino acid; epitope mapping; library; mAb, monoclonal Ab; phage display; site-directed mutagenesis