A Versatile Approach for the Site-Specific Modification of Recombinant Antibodies Using a Combination of Enzyme-Mediated Bioconjugation and Click Chemistry Academic Article uri icon

abstract

  • A unique two-step modular system for site-specific antibody modification and conjugation is reported. The first step of this approach uses enzymatic bioconjugation with the transpeptidase Sortase A for incorporation of strained cyclooctyne functional groups. The second step of this modular approach involves the azide-alkyne cycloaddition click reaction. The versatility of the two-step approach has been exemplified by the selective incorporation of fluorescent dyes and a positron-emitting copper-64 radiotracer for fluorescence and positron-emission tomography imaging of activated platelets, platelet aggregates, and thrombi, respectively. This flexible and versatile approach could be readily adapted to incorporate a large array of tailor-made functional groups using reliable click chemistry whilst preserving the activity of the antibody or other sensitive biological macromolecules.

authors

  • Alt, Karen
  • Paterson, Brett M
  • Westein, Erik
  • Rudd, Stacey E
  • Poniger, Stan S
  • Jagdale, Shweta
  • Ardipradja, Katie
  • Connell, Timothy U
  • Krippner, Guy Y
  • Nair, Ashish KN
  • Wang, Xiaowei
  • Tochon-Danguy, Henri J
  • Donnelly, Paul S
  • Peter, Karlheinz
  • Hagemeyer, Christoph E

publication date

  • 2015