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Dual, Site‐Specific Modification of Antibodies by Using Solid‐Phase Immobilized Microbial Transglutaminase
Author(s) -
Spycher Philipp R.,
Amann Christian A.,
Wehrmüller Jöri E.,
Hurwitz David R.,
Kreis Olivier,
Messmer Daniel,
Ritler Andreas,
Küchler Andreas,
Blanc Alain,
Béhé Martin,
Walde Peter,
Schibli Roger
Publication year - 2017
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201700188
Subject(s) - tissue transglutaminase , bifunctional , chemistry , fluorescence , bioconjugation , antibody , conjugated system , combinatorial chemistry , chelation , bispecific antibody , biochemistry , polymer , enzyme , monoclonal antibody , organic chemistry , catalysis , biology , physics , quantum mechanics , immunology
Abstract Microbial transglutaminase (MTG) was stably solid‐phase immobilized on glass microbeads by using a second‐generation dendronized polymer. Immobilized MTG enabled the efficient generation of site‐specifically conjugated proteins, including antibody fragments, as well as whole antibodies through distinct glutamines and, unprecedentedly, also through lysines with various bifunctional substrates with defined stoichiometries. With this method, we generated dual, site‐specifically modified antibodies comprising a fluorescent probe and a metal chelator for radiolabeling—a strategy anticipated to design antibodies for imaging and simultaneous therapy. Furthermore, we provide evidence that immobilized MTG features higher siteselectivity than soluble MTG.