Construction of di-scFv through a trivalent alkyne–azide 1,3-dipolar cycloaddition
Literature Information
Arutselvan Natarajan, Wenjun Du, Cheng-Yi Xiong, Gerald L. DeNardo, Sally J. DeNardo, Jacquelyn Gervay-Hague
Heterofunctional azide and alkyne PEG-linkers have been synthesized and site specifically conjugated to scFv via a reactive thiol functionality; two scFv were coupled by copper catalyzed 1,3-dipolar cycloaddition to make divalent scFv (di-scFv) with an inter-scFv distance defined to provide divalent binding; antigen binding was maintained for the di-scFv construct and increased several times compared to that of the parent scFv; the cycloaddition reaction reported herein represents an important ligation strategy to covalently link macromolecular proteins and retain sensitive structural conformations.
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