Epitope Mapping for a Preclinical Bevacizumab (Avastin) Biosimilar on an Extended Construct of Vascular Endothelial Growth Factor A Using Millisecond Hydrogen-Deuterium Exchange Mass Spectrometry

Biochemistry. 2020 Aug 4;59(30):2776-2781. doi: 10.1021/acs.biochem.0c00308. Epub 2020 Jul 21.

Abstract

The success of bevacizumab (Avastin), a monoclonal antibody (mAb) anticancer drug targeting vascular endothelial growth factor A (VEGF-A), has motivated the development of biosimilars. Establishing target epitope similarity using epitope mapping is a critical step in preclinical mAb biosimilar development. Here we use time-resolved electrospray ionization hydrogen-deuterium exchange (HDX) mass spectrometry to rapidly compare the epitopes of commercial Avastin and a biosimilar in preclinical development (ApoBev) on an extended construct of VEGF-A. The Avastin and ApoBev epitopes determined in our experiments agree with each other and with the known epitope derived from the Avastin Fab domain/truncated VEGF co-crystal structure. However, subtly different allosteric effects observed exclusively at short (millisecond) HDX labeling times may reflect a slightly different binding mode for ApoBev.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bevacizumab / immunology*
  • Biosimilar Pharmaceuticals / pharmacology*
  • Epitope Mapping*
  • Humans
  • Hydrogen Deuterium Exchange-Mass Spectrometry*
  • Kinetics
  • Microfluidics
  • Models, Molecular
  • Vascular Endothelial Growth Factor A / chemistry*

Substances

  • Biosimilar Pharmaceuticals
  • Vascular Endothelial Growth Factor A
  • Bevacizumab