Vu Bang K, Walsh Joseph D, Dimitrov Dimiter S, Ishima Rieko
NCI-Frederick, National Institutes of Health, Frederick, MD, USA.
Methods Mol Biol. 2009;525:533-43, xv. doi: 10.1007/978-1-59745-554-1_29.
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.
了解抗体的局部动力学信息对于评估稳定性、合理设计变体以及阐明表位结合位点的构象紊乱至关重要。此类信息对于更好地理解抗原识别也可能有用。核磁共振（NMR）可通过弛豫测量在原子水平表征蛋白质的局部动力学。由于NMR谱的复杂性，这种方法的广泛应用仅限于小蛋白质分子，例如抗体结构域和一些单链抗体片段（scFv）。在此，我们描述了一种用于利用NMR研究溶液中抗体结构域动力学的方案。我们介绍了用于NMR研究的蛋白质制备、NMR样品优化、信号归属和动力学实验。
