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Alt K.,Vascular Biotechnology | Paterson B.M.,University of Melbourne | Westein E.,Atherothrombosis and Vascular Biology | Rudd S.E.,University of Melbourne | And 11 more authors.
Angewandte Chemie - International Edition | Year: 2015

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. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Source


Alt K.,Vascular Biotechnology | Paterson B.M.,University of Melbourne | Westein E.,Atherothrombosis and Vascular Biology | Rudd S.E.,University of Melbourne | And 9 more authors.
Angewandte Chemie - International Edition | Year: 2015

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. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Source

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