Abstract
Information on local dynamics of antibodies is important to evaluate stability, to rationally design variants, and to clarify conformational disorders at the epitope binding sites. Such information may also be useful for improved understanding of antigen recognition. NMR can be used for characterization of local protein dynamics at the atomic level through relaxation measurements. Due to the complexity of the NMR spectra, an extensive use of this method is limited to small protein molecules, for example, antibody domains and some scFv. Here, we describe a protocol that was used to study the dynamics of an antibody domain in solution using NMR. We describe protein preparation for NMR studies, NMR sample optimization, signal assignments, and dynamics experiments.
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Acknowledgments
The authors would like to thank Dr. John M. Louis for discussions, and Dr. Stefan Bagby for critical reading of the manuscript. This project was supported by University of Pittsburgh, the NIH Intramural AIDS Targeted Antiviral Program (IATAP), and the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research.
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Vu, B.K., Walsh, J., Dimitrov, D.S., Ishima, R. (2009). Dynamics of Antibody Domains Studied by Solution NMR. In: Dimitrov, A. (eds) Therapeutic Antibodies. Methods in Molecular Biology™, vol 525. Humana Press. https://doi.org/10.1007/978-1-59745-554-1_29
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DOI: https://doi.org/10.1007/978-1-59745-554-1_29
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