Conformational analysis of recombinant monoclonal antibodies with hydrogen/deuterium exchange mass spectrometry.
Conformational analysis of recombinant monoclonal antibodies with hydrogen/deuterium exchange mass spectrometry.
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DOI：
10.1007/978-1-62703-327-5_17
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发表时间：
2013-01-01
期刊：
影响因子：
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通讯作者：
Engen, John R
中科院分区：
文献类型：
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作者：
Houde, Damian;Engen, John R
了解抗体的构象，特别是那些具有治疗价值的抗体，是非常有趣的。目前用于探测蛋白质构象的许多分析方法在抗体分析中面临问题，这是由于抗体本身的性质或由于方法的局限性。最近用于抗体构象分析的一种方法是氢/氘交换质谱法（H/DX MS）。H/DX MS可用于探测溶液中蛋白质的构象和动力学，需要少量样品，与许多缓冲系统兼容，并提供肽级分辨率。H/DX MS应用于免疫球蛋白γ 1（IgG 1）重组单克隆抗体可以提供有关IgG 1构象、动力学和蛋白质修饰导致的构象变化、储存条件变化、纯化程序、制剂和许多其他参数的信息。在这篇文章中，我们提供了一个全面的H/DX MS协议的抗体分析。
Understanding the conformation of antibodies, especially those of therapeutic value, is of great interest. Many of the current analytical methods used to probe protein conformation face issues in the analysis of antibodies, either due to the nature of the antibody itself or due to the limitations of the method. One method that has recently been utilized for conformational analysis of antibodies is hydrogen/deuterium exchange mass spectrometry (H/DX MS). H/DX MS can be used to probe the conformation and dynamics of proteins in solution, requires small sample quantities, is compatible with many buffer systems, and provides peptide-level resolution. The application of H/DX MS to immunoglobulin gamma 1 (IgG1) recombinant monoclonal antibodies can provide information about IgG1 conformation, dynamics, and changes to conformation as a result of protein modification(s), changes in storage conditions, purification procedures, formulation, and many other parameters. In this article we provide a comprehensive H/DX MS protocol for the analysis of an antibody.