Epitope and Paratope Mapping Reveals Temperature-Dependent Alterations in the Dengue-Antibody Interface

Structure. 2017 Sep 5;25(9):1391-1402.e3. doi: 10.1016/j.str.2017.07.007. Epub 2017 Aug 17.


Uncovering mechanisms of antibody-mediated neutralization for viral infections requires epitope and paratope mapping in the context of whole viral particle interactions with the antibody in solution. In this study, we use amide hydrogen/deuterium exchange mass spectrometry to describe the interface of a dengue virus-neutralizing antibody, 2D22, with its target epitope. 2D22 binds specifically to DENV2, a serotype showing strain-specific structural expansion at human host physiological temperatures of 37°C. Our results identify the heavy chain of 2D22 to be the primary determinant for binding DENV2. Temperature-mediated expansion alters the mode of interaction of 2D22 binding. Importantly, 2D22 interferes with the viral expansion process and offers a basis for its neutralization mechanism. The relative magnitude of deuterium exchange protection upon antibody binding across the various epitope loci allows a deconstruction of the antibody-viral interface in host-specific environments and offers a robust approach for targeted antibody engineering.

Keywords: HDXMS; antibodies; dengue virus serotype 2; epitopes; neutralizing antibody-whole dengue virus particle interface mapping; paratopes; structural mass spectrometry; temperature-dependent expansion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Neutralizing / chemistry*
  • Antibodies, Neutralizing / metabolism*
  • Antibodies, Viral / chemistry
  • Antibodies, Viral / metabolism
  • Binding Sites, Antibody
  • Dengue Virus / chemistry
  • Dengue Virus / immunology*
  • Dengue Virus / metabolism
  • Deuterium Exchange Measurement
  • Epitope Mapping
  • Epitopes / chemistry
  • Humans
  • Mass Spectrometry
  • Models, Molecular
  • Protein Binding
  • Protein Conformation
  • Temperature
  • Viral Envelope Proteins / chemistry*
  • Viral Envelope Proteins / metabolism*


  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Epitopes
  • Viral Envelope Proteins