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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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Abstract

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.

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Acknowledgements

The authors thank Drs. Olafur Gudmundsson, Arvind Rajpal, Lois Lehman-McKeeman and Bruce Car from Bristol-Myers Squibb for their support of this project.

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Authors and Affiliations

  1. Pharmaceutical Candidate Optimization, Research and Development, Bristol-Myers Squibb Company, Princeton, NJ, 08543, USA

    Richard Y.-C. Huang, Yun Wang & Guodong Chen

  2. Discovery Biology, Research and Development, Bristol-Myers Squibb Company, Redwood City, CA, 94063, USA

    Michelle Kuhne

  3. Protein Engineering, Research and Development, Bristol-Myers Squibb Company, Redwood City, CA, 94063, USA

    Shrikant Deshpande, Vangipuram Rangan & Mohan Srinivasan

Authors
  1. Richard Y.-C. Huang
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  2. Michelle Kuhne
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  5. Mohan Srinivasan
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  6. Yun Wang
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Correspondence to Richard Y.-C. Huang or Guodong Chen.

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Huang, R.YC., Kuhne, M., Deshpande, S. et al. 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. Anal Bioanal Chem 412, 1693–1700 (2020). https://doi.org/10.1007/s00216-020-02409-x

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  • DOI: https://doi.org/10.1007/s00216-020-02409-x

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