Summary
The aim of this chapter is to provide a strategy for mapping linear antibody epitopes of protein antigens in order to discover candidates for vaccines or diagnostic tests. A set of overlapping peptides was designed and synthesised based upon a known amino acid sequence of the target protein, virulence-associated protein A (VapA) 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 micro titre plates that had been pre-coated with NeutrAvidin™. A linear B-cell epitope was identified by a universal recognition of sera to the synthetic peptides which corresponds to a particular fragment of the VapA protein.
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References
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Acknowledgements
We thank Stuart Rodda of Mimotopes for his advice and helpful discussion. We are grateful to Glenn Browning from University of Melbourne for kindly providing the sera. We also thank Tuck Weng Kok and the staff of the Serology Unit of the Institute of Medical and Veterinary Science for the use of equipment and facilities. This work was supported by the Rural Industries Research and Development Corporation (RIRDC) – Horse Programme and Vet Biotechnology Ltd, Adelaide, South Australia.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Heuzenroeder, M.W., Barton, M.D., Vanniasinkam, T., Phumoonna, T. (2009). Linear B-Cell Epitope Mapping Using Enzyme-Linked Immunosorbent Assay for Libraries of Overlapping Synthetic Peptides. In: Schutkowski, M., Reineke, U. (eds) Epitope Mapping Protocols. Methods in Molecular Biology™, vol 524. Humana Press. https://doi.org/10.1007/978-1-59745-450-6_10
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DOI: https://doi.org/10.1007/978-1-59745-450-6_10
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