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Epitope Mapping Protocols pp 119–134Cite as

Epitope Mapping by Differential Chemical Modification of Antigens

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 524))

Summary

Matrix-assisted laser desorption ionization or electrospray ionization mass spectrometry combined with differential chemical modification have proven to be versatile tools for epitope mapping as well as for studying diverse protein–protein and protein–ligand interactions. Characterization of a discontinuous or a conformational epitope on an antigen demands the ability to map the three-dimensional protein surface along with the interface of two interacting proteins. Classical methods of differentially derivatizing amino acid residues have been successfully merged with highly sensitive and highly accurate mass spectrometric techniques to rapidly profile the three-dimensional protein surface and determine the surface accessibility of specific amino acid residues. Here we discuss the use of mass spectrometry to characterize discontinuous or conformational epitopes by studying antigen–antibody interactions. The steps involved in epitope mapping approaches using differential chemical modification and H/D exchange on the antigen are discussed in detail, with particular emphasis on the experimental protocols.

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Acknowledgments

We thank Dr. Leesa J. Deterding and Dr. Jason G. Williams for helpful discussions. This work was supported by Intramural Research program, National Institute of Environmental Health Sciences, NIH.

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Authors and Affiliations

  1. Laboratory of Structural Biology, National Institute of Environmental Health Sciences, NIH, DHHS, 111 T.W. Alexander Drive, PO Box 12233, Research Triangle Park, NC, 27709, USA

    Suraj Dhungana, Michael B. Fessler & Kenneth B. Tomer

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  1. Suraj Dhungana
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  2. Michael B. Fessler
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  3. Kenneth B. Tomer
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Correspondence to Kenneth B. Tomer .

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    © 2009 Humana Press, a part of Springer Science+Business Media, LLC

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    Dhungana, S., Fessler, M.B., Tomer, K.B. (2009). Epitope Mapping by Differential Chemical Modification of Antigens. 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_9

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    • DOI: https://doi.org/10.1007/978-1-59745-450-6_9

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    • Publisher Name: Humana Press

    • Print ISBN: 978-1-934115-17-6

    • Online ISBN: 978-1-59745-450-6

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