Construction of di-scFv through a trivalent alkyne–azide 1,3-dipolar cycloaddition

Literature Information

Publication Date 2006-11-28
DOI 10.1039/B611636A
Impact Factor 6.222
Authors

Arutselvan Natarajan, Wenjun Du, Cheng-Yi Xiong, Gerald L. DeNardo, Sally J. DeNardo, Jacquelyn Gervay-Hague


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Abstract

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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