title: Peptide microarrays for the characterization of antigenic regions of human chromogranin A.
abstract: Microarraying peptides is a powerful proteomics technique for studying molecular recognition events. Since peptides have small molecular mass, they are not easily accessible when adsorbed onto solid supports. Moreover, peptides can lack a well-defined three-dimensional structure, and therefore a correct orientation is essential to promote the interaction with their target. In this work, we investigated the suitability as a peptide array substrate of a glass slide coated with a copolymer of N,N-dimethylacrylamide, N,N-acryloyloxysuccinimide, and [3-(methacryloyl-oxy)propyl]trimethoxysilyl. This polymeric surface was used as substrate for peptides in the characterization of linear antigenic sites of human chromogranin A, a useful tissue and serum marker for neuroendocrine tumors and a precursor of many biologically active peptides. The microarray support provided sufficient accessibility of the ligand, with no need for a spacer, as the polymer chains prevent interaction of immobilized peptides with substrate. In addition, the polymeric surface constitutes an aqueous micro-environment in which linear epitopes are freely exposed despite peptide random orientation. The results reported in this article are in accordance with those obtained in conventional ELISA assays using biotinylated and non-biotinylated peptides.
authors.affiliation: Istituto di Chimica del Riconoscimento Molecolare, Milan, Italy.
authors.forename: Marcella
authors.initials: M
authors.lastname: Chiari
authors.forename: Marina
authors.initials: M
authors.lastname: Cretich
authors.forename: Angelo
authors.initials: A
authors.lastname: Corti
authors.forename: Francesco
authors.initials: F
authors.lastname: Damin
authors.forename: Giovanna
authors.initials: G
authors.lastname: Pirri
authors.forename: Renato
authors.initials: R
authors.lastname: Longhi
mesh_terms: D000595:Amino Acid Sequence; D000941:Antigens / Q000737:chemistry* / Q000276:immunology; D053379:Chromogranin A; D002864:Chromogranins / Q000737:chemistry* / Q000276:immunology; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000276:immunology; D040081:Protein Array Analysis
chemical_list: D000941:Antigens; C506150:CHGA protein, human; D053379:Chromogranin A; D002864:Chromogranins; D000939:Epitopes; D010455:Peptides