title: Helical peptide arrays for lead identification and interaction site mapping.
abstract: Libraries composed of linear and cyclic peptides cannot fully represent the higher order structures of most antigenic sites. To map the binding site of ligands or antibodies, a larger part of the three-dimensional space should be sampled. Because parallel synthesis of large arrays of peptides on hydrogels is restricted to relatively small peptides, a simple and robust homodimeric helical system was chosen for antigen presentation. First, it was established in an heterodimeric system that the 26-mer peptide could be synthesized and that the helical coiled-coil peptides interact in the hydrogel in a predictable manner. Next, libraries of homodimeric coiled coils were synthesized into which the epitope was grafted. Using dedicated helical dimeric and trimeric coiled-coil libraries, the epitopes of two anti-HIV-1 gp41 monoclonal antibodies known to interact with helical structures were mapped at high resolution. These mappings precisely reflect existing X-ray data, and the arrays can be applied to lead identification, epitope mapping, and systematic analysis of amino acid contribution to coiled-coil systems.
authors.affiliation: Pepscan Therapeutics, 8243 RC Lelystad, The Netherlands. hans.langedijk@crucell.com
authors.forename: Johannes P M
authors.initials: JP
authors.lastname: Langedijk
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Zekveld
authors.forename: Mariska
authors.initials: M
authors.lastname: Ruiter
authors.forename: Davide
authors.initials: D
authors.lastname: Corti
authors.forename: Jaap W
authors.initials: JW
authors.lastname: Back
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001710:Biotin / Q000378:metabolism; D018604:Epitope Mapping; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D017433:Protein Structure, Secondary; D012995:Solubility
chemical_list: D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D010455:Peptides; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D001710:Biotin