Generating site-specifically modified proteins via a versatile and stable nucleophilic carbon ligation

Chem Biol. 2015 Feb 19;22(2):293-8. doi: 10.1016/j.chembiol.2014.11.019. Epub 2015 Jan 22.

Abstract

There is a need for facile chemistries that allow for chemo- and regioselectivity in bioconjugation reactions. To address this need, we are pioneering site-specific bioconjugation methods that use formylglycine as a bioorthogonal handle on a protein surface. Here we introduce aldehyde-specific bioconjugation chemistry, the trapped-Knoevenagel ligation. The speed and stability of the trapped-Knoevenagel ligation further advances the repertoire of aldehyde-based bioconjugations and expands the toolbox for site-specific protein modifications. The trapped-Knoevenagel ligation reaction can be run at near neutral pH in the absence of catalysts to produce conjugates that are stable under physiological conditions. Using this new ligation, we generated an antibody-drug conjugate that demonstrates excellent efficacy in vitro and in vivo.

Publication types

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

MeSH terms

  • Antineoplastic Agents / chemical synthesis
  • Antineoplastic Agents / chemistry
  • Antineoplastic Agents / toxicity
  • Brentuximab Vedotin
  • Carbon / chemistry*
  • Catalysis
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Humans
  • Hydrogen-Ion Concentration
  • Immunoconjugates / chemistry
  • Proteins / chemistry*
  • Pyrazoles / chemistry
  • Trastuzumab / chemistry

Substances

  • Antineoplastic Agents
  • Immunoconjugates
  • Proteins
  • Pyrazoles
  • Carbon
  • Brentuximab Vedotin
  • Trastuzumab