TY - JOUR
T1 - Enzyme-mediated site-specific bioconjugation of metal complexes to proteins: Sortase-mediated coupling of copper-64 to a single-chain antibody
AU - Paterson, Brett M
AU - Alt, Karen Maria
AU - Jeffery, Charmaine M
AU - Price, Roger I
AU - Jagdale, Schweta
AU - Williams, Charlotte Claire
AU - Rigby, Sheena
AU - Peter, Karlheinz
AU - Hagemeyer, Christoph Eugen
AU - Donnelly, Paul Stephen
PY - 2014
Y1 - 2014
N2 - The enzyme-mediated site-specific bioconjugation of a radioactive metal complex to a single-chain antibody using the transpeptidase sortase A is reported. Cage amine sarcophagine ligands that were designed to function as substrates for the sortase A mediated bioconjugation to antibodies were synthesized and enzymatically conjugated to a single-chain variable fragment. The antibody fragment scFvanti-LIBS targets ligand-induced binding sites (LIBS) on the glycoprotein receptor GPIIb/IIIa, which is present on activated platelets. The immunoconjugates were radiolabeled with the positron-emitting isotope 64Cu. The new radiolabeled conjugates were shown to bind selectively to activated platelets. The diagnostic potential of the most promising conjugate was demonstrated in an in vivo model of carotid artery thrombosis using positron emission tomography. This approach gives homogeneous products through site-specific enzyme-mediated conjugation and should be broadly applicable to other metal complexes and proteins.
AB - The enzyme-mediated site-specific bioconjugation of a radioactive metal complex to a single-chain antibody using the transpeptidase sortase A is reported. Cage amine sarcophagine ligands that were designed to function as substrates for the sortase A mediated bioconjugation to antibodies were synthesized and enzymatically conjugated to a single-chain variable fragment. The antibody fragment scFvanti-LIBS targets ligand-induced binding sites (LIBS) on the glycoprotein receptor GPIIb/IIIa, which is present on activated platelets. The immunoconjugates were radiolabeled with the positron-emitting isotope 64Cu. The new radiolabeled conjugates were shown to bind selectively to activated platelets. The diagnostic potential of the most promising conjugate was demonstrated in an in vivo model of carotid artery thrombosis using positron emission tomography. This approach gives homogeneous products through site-specific enzyme-mediated conjugation and should be broadly applicable to other metal complexes and proteins.
UR - http://onlinelibrary.wiley.com/doi/10.1002/anie.201402613/abstract
U2 - 10.1002/anie.201402613
DO - 10.1002/anie.201402613
M3 - Article
SN - 1433-7851
VL - 53
SP - 6115
EP - 6119
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 24
ER -