Epitope mapping of human chromogranin A by peptide microarrays.
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.
Cretich
Marina
M
Istituto di Chimica del Riconoscimento Molecolare (ICRM)-C.N.R. Milano, Milano, Italy.
Longhi
Renato
R
Corti
Angelo
A
Damin
Francesco
F
Di Carlo
Gabriele
G
Sedini
Valentina
V
Chiari
Marcella
M
D053379:Chromogranin A / Q000737:chemistry* / Q000276:immunology; D018604:Epitope Mapping / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000494:pharmacology; D006801:Humans; D008954:Models, Biological; D010455:Peptides / Q000032:analysis / Q000138:chemical synthesis; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*
D053379:Chromogranin A; D005456:Fluorescent Dyes; D010455:Peptides