The aim of this chapter is to provide a strategy for mapping linear antibody epitopes of protein antigens in order to find vaccine or diagnostic test candidates. A set of overlapping peptides was designed and synthesised based upon a known amino acid sequence of the target protein;, VapA (virulence-associated protein A) of the bacterium Rhodococcus equi, an important pulmonary pathogen in foals. The peptides were biotinylated and used in an ELISA to screen immune sera from foals. These biotinylated peptides were coated directly onto microtitre plates which had been pre-coated with NeutrAvidin. A linear B-cell epitope was identified by a universal recognition of sera to the synthetic peptides which corresponded to a particular fragment of the VapA protein.
|Title of host publication||Methods in Molecular Biology|
|Subtitle of host publication||Epitope mapping protocols|
|Editors||Ulrich Reineke, Mike Schutkowski|
|Place of Publication||New Jersey, USA|
|Number of pages||8|
|Publication status||Published - 2008|