Construction of di-scFv through a trivalent alkyne-azide 1,3-dipolar cycloaddition
Construction of di-scFv through a trivalent alkyne-azide 1,3-dipolar cycloaddition
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DOI：
10.1039/b611636a
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发表时间：
2007-01-01
影响因子：
4.9
通讯作者：
Gervay-Hague, Jacquelyn
中科院分区：
文献类型：
--
作者：
Natarajan, Arutselvan;Du, Wenjun;Gervay-Hague, Jacquelyn
已经合成了异功能叠氮化物和炔PEG-接头，并通过反应性硫醇官能团与scFv位点特异性缀合;两个scFv通过铜催化的1，3-偶极环加成偶联，以制备二价scFv（discFv），其具有限定的scFv间距离以提供二价结合;双-scFv构建体保持抗原结合，并且与亲本scFv相比增加数倍;本文报道的环加成反应代表了共价连接大分子蛋白质并保持敏感的结构构象的重要连接策略。
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 (discFv) 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.