A versatile approach for the site-specific modification of recombinant antibodies using a combination of enzyme-mediated bioconjugation and click chemistry

Karen Maria Alt, Brett M Paterson, Erik Westein, Stacey Rudd, Stan Poniger, Schweta Jagdale, Katie Ardipradja, Timothy U Connell, Guy Yeoman Krippner, Ashish Kumar Narayan Nair, Xiaowei Wang, Henri Tochon-Danguy, Paul Stephen Donnelly, Karlheinz Peter, Christoph Eugen Hagemeyer

Research output: Contribution to journalArticleResearchpeer-review

27 Citations (Scopus)

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. A 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 site-specific incorporation of strained cyclooctyne functional groups into antibodies. The second step of this modular approach involves the copper-free azide-alkyne cycloaddition click reaction.
Original languageEnglish
Pages (from-to)7515 - 7519
Number of pages5
JournalAngewandte Chemie - International Edition
Volume54
Issue number26
DOIs
Publication statusPublished - 2015
Externally publishedYes

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