天然 MS 和 ECD 对 Fab-抗原复合物的分析有助于 X 射线衍射的结晶。
Native MS and ECD Characterization of a Fab-Antigen Complex May Facilitate Crystallization for X-ray Diffraction.
机构信息
Department of Chemistry, Washington University in St. Louis, One Brookings Dr., St. Louis, MO, 63130, USA.
Analytical Research and Development, Pfizer Inc., Chesterfield, MO, 63017, USA.
出版信息
J Am Soc Mass Spectrom. 2016 Jul;27(7):1139-42. doi: 10.1007/s13361-016-1398-9. Epub 2016 Apr 21.
Abstract
Native mass spectrometry (MS) and top-down electron-capture dissociation (ECD) combine as a powerful approach for characterizing large proteins and protein assemblies. Here, we report their use to study an antibody Fab (Fab-1)-VEGF complex in its near-native state. Native ESI with analysis by FTICR mass spectrometry confirms that VEGF is a dimer in solution and that its complex with Fab-1 has a binding stoichiometry of 2:2. Applying combinations of collisionally activated dissociation (CAD), ECD, and infrared multiphoton dissociation (IRMPD) allows identification of flexible regions of the complex, potentially serving as a guide for crystallization and X-ray diffraction analysis. Graphical Abstract ᅟ.
摘要
天然质谱（MS）和自上而下的电子俘获解离（ECD）相结合，是一种用于表征大型蛋白质和蛋白质组装体的强大方法。在这里，我们报告了它们在研究其近天然状态下的抗体 Fab（Fab-1）-VEGF 复合物中的应用。天然 ESI 与傅立叶变换离子回旋共振质谱分析相结合，证实 VEGF 在溶液中为二聚体，并且其与 Fab-1 的复合物具有 2:2 的结合计量比。应用碰撞激活解离（CAD）、ECD 和红外多光子解离（IRMPD）的组合，可以鉴定复合物的柔性区域，这可能为结晶和 X 射线衍射分析提供指导。
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