TY - JOUR
T1 - Peptide microarrays for the characterization of antigenic regions of human chromogranin A
AU - Chiari, Marcella
AU - Cretich, Marina
AU - Corti, Angelo
AU - Damin, Francesco
AU - Pirri, Giovanna
AU - Longhi, Renato
PY - 2005/9
Y1 - 2005/9
N2 - 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.
AB - 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.
KW - Human chromogranin A
KW - Peptide microarray
KW - Polymer coating
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U2 - 10.1002/pmic.200401216
DO - 10.1002/pmic.200401216
M3 - Article
C2 - 16097033
AN - SCOPUS:25844458238
VL - 5
SP - 3600
EP - 3603
JO - Proteomics
JF - Proteomics
SN - 1615-9853
IS - 14
ER -