Mapping binding epitopes of monoclonal antibodies targeting major histocompatibility complex class I chain-related A (MICA) with hydrogen/deuterium exchange and electron-transfer dissociation mass spectrometry
Mapping binding epitopes of monoclonal antibodies targeting major histocompatibility complex class I chain-related A (MICA) with hydrogen/deuterium exchange and electron-transfer dissociation mass spectrometry
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DOI：
10.1007/s00216-020-02409-x
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发表时间：
2020-03-01
影响因子：
4.3
通讯作者：
Chen，Guodong
中科院分区：
文献类型：
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作者：
Huang，Richard Y. -C.;Kuhne，Michelle;Chen，Guodong
主要组织相容性复合体I类链相关A和B （MICA/B）是细胞表面蛋白，作为自然杀伤细胞受体（NKG2D）的配体，在免疫细胞上表达。预防MICA/B蛋白水解脱落以保持其在细胞表面的完整性已成为免疫肿瘤学的治疗策略。鉴于MICA/B脱落的独特机制，MICA/B和治疗剂相互作用的结构表征在药物发现过程中很重要。在这项研究中，我们描述了氢/氘交换质谱（HDX-MS）在四种靶向MICA的单克隆抗体的快速表位定位研究中的实际应用。HDX-MS之后的电子转移解离允许结合表位的高分辨率细化。这一综合策略首次提供了对MICA在溶液中的构象动力学的分子水平理解，以及这些抗体靶向MICA的独特作用机制。图形抽象
Major histocompatibility complex class I chain-related A and B (MICA/B) are cell-surface proteins that act as ligands to natural killer cell receptors, NKG2D, expressed on immune cells. Prevention of proteolytic shedding of MICA/B to retain their integrity on the cell surface has become a therapeutic strategy in immuno-oncology. Given the unique mechanism of MICA/B shedding, structural characterization of MICA/B and therapeutic agent interaction is important in the drug discovery process. In this study, we describe the practical utility of hydrogen/deuterium exchange mass spectrometry (HDX-MS) in epitope mapping studies of a cohort of four monoclonal antibodies targeting MICA in a rapid manner. HDX-MS followed by electron-transfer dissociation allows high-resolution refinement of binding epitopes. This integrated strategy offers, for the first time, molecular-level understanding of MICA’s conformational dynamics in solution as well as the unique mechanism of actions of these antibodies in targeting MICA.Graphical abstract