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10. Epitope Mapping of Human Chromogranin A by Peptide Microarrays
Abstract
In this chapter we report on the characterization of linear antigenic sites of human chromogranin A (CgA), a useful tissue and serum marker for neuroendocrine tumours and a precursor of many biologically active peptides. The epitope mapping of CgA has been carried out by peptide microarrays on glass slides coated by a copolymer of N,N-dimethylacrylamide (DMA), N,N-acryloyloxysuccinimide (NAS) and [3-(methacryloyl-oxy) propyl] trimethoxysilyl (MAPS). 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, despite peptide random orientation, linear epitopes are freely exposed. The results reported are in accordance with those obtained in conventional ELISA assays using biotinylated and non-biotinylated peptides.
Affiliation(s): (1) Istituto di Chimica del Riconoscimento Molecolare (ICRM) – C.N.R. Milano, Milano, Italy
(2) Immunobiotechnology Unit, Department of Biological and Technological Research, San Raffaele H Scientific Institute, Milano, Italy
Series: Methods in Molecular Biology  |  Volume: 570  |  Pub. Date: Aug-01-2009  |  Page Range: 221-232  |  DOI: 10.1007/978-1-60327-394-7_10
Subject:  Protein Science
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