Epitope mapping of human chromogranin A by peptide microarrays

Methods Mol Biol. 2009:570:221-32. doi: 10.1007/978-1-60327-394-7_10.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Chromogranin A / chemistry*
  • Chromogranin A / immunology
  • Epitope Mapping / instrumentation
  • Epitope Mapping / methods*
  • Fluorescent Dyes / pharmacology
  • Humans
  • Models, Biological
  • Peptides / analysis
  • Peptides / chemical synthesis
  • Protein Array Analysis / instrumentation
  • Protein Array Analysis / methods*

Substances

  • Chromogranin A
  • Fluorescent Dyes
  • Peptides