Linear B-Cell Epitope Mapping Using Libraries of Overlapping Synthetic Peptides based Enzyme-Linked Immunosorbent Assay

Thirumahal Vanniasinkam, Michael Heuzenroeder, Mary Barton, Tongted Phumoonna

Research output: Book chapter/Published conference paperChapter

8 Citations (Scopus)
12 Downloads (Pure)

Abstract

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.
Original languageEnglish
Title of host publicationMethods in Molecular Biology
Subtitle of host publicationEpitope mapping protocols
EditorsUlrich Reineke, Mike Schutkowski
Place of PublicationNew Jersey, USA
PublisherHumana Press
Chapter10
Pages137-144
Number of pages8
Volume524
Edition2nd
ISBN (Electronic)9781597454506
ISBN (Print)9781934115176
DOIs
Publication statusPublished - 2008

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