title: Helical epitopes determined by low-stringency antibody screening of a combinatorial peptide library.
abstract: Combinatorial phage display peptide libraries are routinely used to map epitopes of specific monoclonal antibodies. In this study we illustrate that these libraries can be used in the analysis of protein structure. By screening libraries at low stringency, a collection of phages can be obtained. These are characterized by the fact that they are recognized by a given monoclonal antibody yet with various affinities. Comparing the random peptides of these phages indicates the common essential residues necessary for antibody recognition. Aligning the inserts based on the detected homology has revealed structural motifs that correspond to secondary protein structures. The envelope protein of HIV-1 has been studied using this approach. A combinatorial phage display library containing a 20 mer random peptide in protein III of the filamentous phage fd-tet has been used to analyze two different monoclonal antibodies directed against gp120. Our results provide experimental evidence that indicate that the C1 domain of gp120 contains an alpha helix.
authors.affiliation: Department of Cell Research and Immunology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Israel.
authors.forename: B
authors.initials: B
authors.lastname: Stern
authors.forename: G
authors.initials: G
authors.lastname: Denisova
authors.forename: D
authors.initials: D
authors.lastname: Buyaner
authors.forename: D
authors.initials: D
authors.lastname: Raviv
authors.forename: J M
authors.initials: JM
authors.lastname: Gershoni
mesh_terms: D000595:Amino Acid Sequence / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000915:Antibody Affinity; D003090:Coliphages / Q000235:genetics; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D005822:Genetic Vectors; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D015497:HIV-1 / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D015699:HIV Envelope Protein gp120; D010455:Peptides