Substrate Design Enables Heterobifunctional, Dual "Click" Antibody Modification via Microbial Transglutaminase

Bioconjug Chem. 2019 Sep 18;30(9):2452-2457. doi: 10.1021/acs.bioconjchem.9b00522. Epub 2019 Aug 21.

Abstract

Site-specific modification of native antibodies has proven advantageous, as it enhances the properties of antibody-based bioconjugates without the need to manipulate the genetic code. However, native antibody modification is typically limited to strategies that introduce a single functional handle. In this work, we addressed this limitation by designing heterobifunctional substrates for microbial transglutaminase (MTG) that contain both azide and methyltetrazine "click" handles. Structure-conjugation relationships for these substrates were evaluated using the Her2-targeted antibody trastuzumab. Förster resonance energy transfer (FRET) was used to demonstrate that these chemical handles are mutually orthogonal. This orthogonality was leveraged for the one-pot synthesis of a bifunctional antibody-drug conjugate (ADC). This ADC, containing a maytansine-derived payload and a hydrophobicity-masking polyethylene glycol (PEG) side chain, demonstrated potent in vitro activity in SKOV3 cells. These studies establish the dual "click" approach as a powerful technique in the toolbox for native antibody modification.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cell Line, Tumor
  • Click Chemistry
  • Cysteine / chemistry
  • HEK293 Cells
  • Humans
  • Immunoconjugates / chemistry*
  • Immunoconjugates / metabolism*
  • Microbiology
  • Oxidation-Reduction
  • Transglutaminases / metabolism*
  • Tryptophan / chemistry

Substances

  • Immunoconjugates
  • Tryptophan
  • Transglutaminases
  • Cysteine