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.
CITATION STYLE
Alt, K., Paterson, B. M., Westein, E., Rudd, S. E., Poniger, S. S., Jagdale, S., … Hagemeyer, C. E. (2015). A versatile approach for the site-specific modification of recombinant antibodies using a combination of enzyme-mediated bioconjugation and click chemistry. Angewandte Chemie - International Edition, 54(26), 7515–7519. https://doi.org/10.1002/anie.201411507
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