Abstract
The region of an antigen that interacts with an antibody is defined as an epitope. For protein antigens, epitopes may involve a single length of the polypeptide chain (sequential or linear epitopes) or may be composed of several widely separated, discrete amino acid sequences that come together in the folded native portion (conformational or discontinuous epitopes) (1). Complete definition of the structure of an epitope can be achieved by X-ray crystallography of antigen-antibody cocrystals, but to date only a limited number of protein epitopes (all of the discontinuous type) have been defined by this method (1,2). These studies, however, have suggested that the epitopes of native protein consist of 15–22 residues with a smaller subset of 5–6 residues contributing most of the binding energy. It is important to note that these critical residues may not be arranged in a linear sequence (1).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Laver, W. G., Air, G. M. Webster, R. G., and Smith-Gill, S. J. (1990) Epitopes on protein antigens: Misconceptions and realities. Cell 61, 553–556.
Davies, D. R. and Cohen, G. H. (1996) Interactions of protein antigens with antibodies. Proc. Natl. Acad. Sci. USA 74, 5463–5467.
DeLisser, H. M., Baldwin, H. S., and Albelda, S. M. (1997) PECAM-1/CD31-a multifunctional vascular cell adhension molecule. Trends Cardiovasc. Med. 8, 203–210.
Yan, H., Pilewski, J. M., Zhang, Q., DeLisser, H. M., Romer, L., and Albelda, S. M. Localization of multiple functional domains on human PECAM-1 (CD31) by monoclonal antibody epitope mapping. Cell Adhesion Commun. 3, 45–66.
Liao, F., Huynh, H. K., Eiro, A., Greene, T., Polizzi, E., and Muller, M. A. (1995) Migration of monocytes across endothelium and passage through extracellular matrix involve seperate molecular domains of PECAM-1. J. Exp. Med. 182, 1337–1343.
Ashman, L. K., Aylett, G. W., Cambareri, A. C., and Cole, S. R. (1991) Different epitopes of the CD31 antigen identified by monoclonal antibodies: cell type-specific patterns of expression. Tissue Antigens 38, 199–207.
Roost, H. P., Haag, A., Burkhart, C., and Zinkernagel, R. M. (1996) Mapping of the dominant neutralizing antigenic site of a virus using infected cells. J. Immunol. Methods 189, 233–242.
Perton, F. G., Dijkema, J. H., Smilda, T., Erikvan Ufflen, B., and Beintema, J. J. (1996) Comparison of three methods for competive binding of monoclonal antibodies. The localization of antigenic sites for monoclonal antibodies on Panulirus interruptus hemocyanin. J. Immunol. Methods 190, 117–125.
Pietu, G., Ribba, A., Cherel, G., Siguret, V., Obert, B., Rouault, C., Ginsburg, D., and Meyer, D. (1994) Epitope mapping of inhibitory monoclonal antibodies to human von Willebrand factor by using recombinant cDNA libraries. Thromb. Haemost. 71, 788–792.
van Zonneveld, A. J. van den Berg, B. M., van Meijer, M., and Pannekoek, H. (1995) Identification of functional interaction sites on proteins using bacteriophage-displayed random epitope libraries. Gene 167, 49–52.
Peterson, G., Song, D., Hugle-Dorr, B., Oldenburg, I., and Bautz, E. K. (1995) Mapping of linear epitopes recognized by monoclonal antibodies with genefragment phage display libraries. Mol. Gen. Gene. 249, 425–431.
Rao Y., Wu, X., Gariepy, J., Rutishauser, U., and Siu, C. (1992) Identification of a peptide sequence involved in homophilic binding in the neural cell adhesion molecule NCAM. J. Cell Biol. 118, 937–949.
Tzartos, S. J. and Remouunds M. S. (1992) Precise epitope mapping of monoclonal antibodies to the cytoplasmic side of the acetycholine receptor a subunit. Eur. J. Biochem. 207, 915–922.
Li, F., Erickson, H. P., James, J. A., Moore K. L. Cummings, R. D., and McEver, R. P. (1996) Visualization of P-selectin glycoprotein ligand-1 as a highly extended molecule and the mapping of protein epitopes for monoclonal antibodies. J. Biol. Chem. 271, 6342–6348.
Ueno, H., Masuko, T., Wang, J., and Hashimoto, Y. (1994) Epitope mapping of bovine serum albumin using monoclonal antibodies coupled with a photoreactive crosslinker. J. Biochem. 115, 1119–1127.
Yuan, J. and Low P. S. (1992) Epitope mapping by a method that requires no amino acid sequence information. Anal. Biochem. 205, 179–182.
Fawcett J., Buckley, C., Holness, C. L., Bird, I. N., Spragg, J. H. Saunders J., Harris, A., and Simmons, D. L. (1995) Mapping the homotypic binding sites in CD31 and the role of CD31 adhesion in the formation of intraendothelial cell contacts. J. Cell. Biol. 128, 1229–1241.
Takada, Y. and Puzon W. (1993) Identification of a regulatory region of integrin β1 subunit using activating and inhibiting antibodies. J. Biol. Chem. 268, 17,597–17,601.
Shih, D., Edleman, J. M., Horwitz, A. F., Grunwald, G. B., and Buck, C. A. (1993) Structure/function analysis of the integrin β1 subunit by epitope mapping. J. Cell. Biol. 122, 1361–1371.
Schiffer, S. G., Hemler, M. E., Lobb, R. R., and Osborn, L. (1995) Molecular mapping of functional antibody binding sites of the alpha 4 integrin. J. Biol. Chem. 270, 14,270–14,273.
Binnerts, M. E. van Kooyk, Y., Edwards, C. P., Champe, M., Presta, L., Bodary, S. C. Figdor, C. G., and Berman, P. W. (1996) Antibodies that selectively inhibit leukocyte-function-associated antigen 1 binding to intracellular adhesion molecule-3 recognize a unique epitope within the CD11a I domain. J. Biol. Chem. 271, 9962–9968.
Tomlinson, M. G., Williams, A. F., and Wright, M. D. (1993) Epitope mapping of anti-rat CD53 monoclonal antibodies. Implications for the membrane orientation of the transmembrane 4 superfamily. Eur. J. Immunol. 23, 136–140.
Sun, W., Cohen, S. A., and Barchi, R. L. (1995) Localization of epitopes for monoclonal antibodies directed against the adult rat skeletal muscle sodium channel (rSkM1) using polymerase chain reaction, fusion proteins and western blotting. Anal. Biochem. 226, 188–191.
Ni, Y., Tominaga, Y., Honda Y., Morimoto, K., Sakamoto, S. and Kawai, A. (1995) Mapping and characterization of a sequential epitope on the rabies virus glycoprotein which is recognized by a neutralizing monoclonal antibody, RG719. Microbiol. Immunol. 39, 693–702.
Bazzoni, G., Shih, D. T., Buck, C. A. and Hemler, M. E. (1995) Monoclonal antibody 9EG7 defines a novel beta 1 intergrin epitope induced by soluble ligand and manganese, but inhibited by calcium. J. Biol. Chem. 270, 25,570–25,577.
Horton, R. M., Cai, Z., Ho, S. N. and Pease, L. R. (1990) Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction. Biotechniques 8, 528–535.
DeLisser, H. M., Yan, H., Newman, P. J., Muller, W. A., Buck, C. A., and Albelda, S. M. (1993) Platelet/endothelial cell adhesion molecule-1 (CD31)-mediated cellular aggregation involves cell surface glycosaminoglycans. J. Biol. Chem. 268, 16,037–16,046.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc.
About this protocol
Cite this protocol
DeLisser, H.M. (1999). Epitope Mapping. In: Dejana, E., Corada, M. (eds) Adhesion Protein Protocols. Methods in Molecular Biology, vol 96. Humana Press. https://doi.org/10.1385/1-59259-258-9:11
Download citation
DOI: https://doi.org/10.1385/1-59259-258-9:11
Publisher Name: Humana Press
Print ISBN: 978-0-89603-417-4
Online ISBN: 978-1-59259-258-6
eBook Packages: Springer Protocols