Dual, Site‐Specific Modification of Antibodies by Using Solid‐Phase Immobilized Microbial Transglutaminase
Dual, Site‐Specific Modification of Antibodies by Using Solid‐Phase Immobilized Microbial Transglutaminase
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DOI：
10.1002/cbic.201700188
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发表时间：
2017-10
期刊：
影响因子：
3.2
通讯作者：
P. Spycher;Christian A Amann;Jöri E. Wehrmüller;D. Hurwitz;Olivier Kreis;D. Messmer;Andreas Ritler;A. Küchler;A. Blanc;M. Béhé;P. Walde;R. Schibli
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文献类型：
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作者：
P. Spycher;Christian A Amann;Jöri E. Wehrmüller;D. Hurwitz;Olivier Kreis;D. Messmer;Andreas Ritler;A. Küchler;A. Blanc;M. Béhé;P. Walde;R. Schibli
采用第二代树枝状聚合物将微生物转氨酶（MTG）稳定地固相固定在玻璃微珠上。固定的MTG使得能够通过不同的谷氨酰胺以及前所未有地还通过赖氨酸与具有限定化学计量的各种双功能底物有效产生位点特异性缀合的蛋白质，包括抗体片段以及完整抗体。通过这种方法，我们产生了双重的，位点特异性修饰的抗体，包括一个荧光探针和一个金属螯合剂的放射性标记-一种策略，预计设计抗体的成像和同时治疗。此外，我们提供的证据表明，固定化MTG功能更高的位点选择性比可溶性MTG。
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.