title: A versatile approach for the site-specific modification of recombinant antibodies using a combination of enzyme-mediated bioconjugation and click chemistry.
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.affiliation: Vascular Biotechnology, Baker IDI, Melbourne (Australia). karen.alt@bakeridi.edu.au.
authors.forename: Karen
authors.initials: K
authors.lastname: Alt
authors.affiliation: School of Chemistry/Bio21 Institute, University of Melbourne (Australia).
authors.forename: Brett M
authors.initials: BM
authors.lastname: Paterson
authors.affiliation: Atherothrombosis and Vascular Biology, Baker IDI, Melbourne (Australia).
authors.forename: Erik
authors.initials: E
authors.lastname: Westein
authors.affiliation: School of Chemistry/Bio21 Institute, University of Melbourne (Australia).
authors.forename: Stacey E
authors.initials: SE
authors.lastname: Rudd
authors.affiliation: Department of Molecular Imaging and Therapy, Austin Health, Melbourne (Australia).
authors.forename: Stan S
authors.initials: SS
authors.lastname: Poniger
authors.affiliation: Vascular Biotechnology, Baker IDI, Melbourne (Australia).
authors.forename: Shweta
authors.initials: S
authors.lastname: Jagdale
authors.affiliation: Vascular Biotechnology, Baker IDI, Melbourne (Australia).
authors.forename: Katie
authors.initials: K
authors.lastname: Ardipradja
authors.affiliation: School of Chemistry/Bio21 Institute, University of Melbourne (Australia).
authors.forename: Timothy U
authors.initials: TU
authors.lastname: Connell
authors.affiliation: Vascular Biotechnology, Baker IDI, Melbourne (Australia).
authors.forename: Guy Y
authors.initials: GY
authors.lastname: Krippner
authors.affiliation: Atherothrombosis and Vascular Biology, Baker IDI, Melbourne (Australia).
authors.forename: Ashish K N
authors.initials: AK
authors.lastname: Nair
authors.affiliation: Atherothrombosis and Vascular Biology, Baker IDI, Melbourne (Australia).
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Molecular Imaging and Therapy, Austin Health, Melbourne (Australia).
authors.forename: Henri J
authors.initials: HJ
authors.lastname: Tochon-Danguy
authors.affiliation: School of Chemistry/Bio21 Institute, University of Melbourne (Australia). pauld@unimelb.edu.au.
authors.forename: Paul S
authors.initials: PS
authors.lastname: Donnelly
authors.affiliation: Atherothrombosis and Vascular Biology, Baker IDI, Melbourne (Australia).
authors.forename: Karlheinz
authors.initials: K
authors.lastname: Peter
authors.affiliation: Vascular Biotechnology, Baker IDI, Melbourne (Australia). christoph.hagemeyer@bakeridi.edu.au.
authors.forename: Christoph E
authors.initials: CE
authors.lastname: Hagemeyer
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D057930:Click Chemistry; D051379:Mice; D015394:Molecular Structure; D049268:Positron-Emission Tomography / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins
keywords: PET imaging; Sortase A; bioconjugation; click chemistry; fluorescence imaging