Abstract
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 20 mer sequence of the VapA protein.
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Acknowledgments
The authors would like to thank Stuart Rodda of Mimotopes for his advice and helpful discussions. We are grateful to Glenn Browning from University of Melbourne for kindly providing the foal sera. We also thank Tuck Weng Kok and staff of Serology Unit of SA Pathology 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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Vanniasinkam, T., Barton, M.D., Das, T.P., Heuzenroeder, M.W. (2018). B-Cell Epitope Mapping Using a Library of Overlapping Synthetic Peptides in an Enzyme-Linked Immunosorbent Assay. In: Rockberg, J., Nilvebrant, J. (eds) Epitope Mapping Protocols. Methods in Molecular Biology, vol 1785. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7841-0_8
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DOI: https://doi.org/10.1007/978-1-4939-7841-0_8
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