This chapter describes a strategy for mapping linear B-cell epitopes of proteins using synthetic biotinylated peptides in an ELISA. A set of overlapping peptides were designed 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 synthesized as biotinylated peptides were coated directly onto micro titer plates which had been pre-coated with NeutrAvidin™ and used to screen sera from foals confirmed to have R. equi disease. A linear B-cell epitope was identified which corresponded to a 20Â mer sequence of the VapA protein.
|Title of host publication||Epitope mapping protocols|
|Editors||Johan Rockberg, Johan Nilvebrant|
|Place of Publication||New York, NY|
|Publisher||Humana Press Inc.|
|Number of pages||8|
|Publication status||Published - 01 May 2018|
|Name||Methods in molecular biology|