Accelerated bioorthogonal conjugation: A practical method for the Ligation of diverse functional molecules to a polyvalent virus scaffold
Accelerated bioorthogonal conjugation: A practical method for the Ligation of diverse functional molecules to a polyvalent virus scaffold
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DOI：
10.1021/bc050147l
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发表时间：
2005-11-01
影响因子：
4.7
通讯作者：
Finn, MG
中科院分区：
文献类型：
--
作者：
Sen Gupta, S;Kuzelka, J;Finn, MG
由于蛋白质的多个未保护的官能团和通常低的浓度，与蛋白质形成共价键变得困难。水溶性磺化红菲咯啉配体（2）用于促进高效的Cu-I介导的叠氮-炔环加成（CuAAC）反应，用于生物相关分子与豇豆花叶病毒（CPMV）的化学选择性连接。连接的底物包括复合糖、肽、聚（环氧乙烷）聚合物和铁载体蛋白转铁蛋白，即使对于先前使用常规配体三（三唑基）胺对叠氮化物-炔偶联具有抗性的病例也取得了常规成功（1）。使用4-6当量的底物足以以60- 85%的产率实现60-115个分子/病毒粒子的载量。虽然它对氧敏感，但Cu（.）2系统使其成为要求苛刻的生物缀合应用的有用工具。
Covalent bond formation to proteins is made difficult by their multiple unprotected functional groups and normally low concentrations. A water-soluble sulfonated bathophenanthroline ligand (2) was used to promote a highly efficient Cu-I-mediated azide-alkyne cycloaddition (CuAAC) reaction for the chemoselective attachment of biologically relevant molecules to cowpea mosaic virus (CPMV). The ligated substrates included complex sugars, peptides, poly(ethylene oxide) polymers, and the iron carrier protein transferrin, with routine success even for cases that were previously resistant to azide-alkyne coupling using the conventional ligand tris(triazolyl)amine (1). The use of 4-6 equiv of substrate was sufficient to achieve loadings of 60-115 molecules/virion in yields of 60-85%. Although it is sensitive to oxygen, the reliably efficient performance of the Cu(.)2 system makes it a useful tool for demanding bioconjugation applications.