Analysis and optimization of copper-catalyzed azide-alkyne cycloaddition for bioconjugation.
Analysis and optimization of copper-catalyzed azide-alkyne cycloaddition for bioconjugation.
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DOI：
10.1002/anie.200905087
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发表时间：
2009
影响因子：
16.6
通讯作者：
Finn, M. G.
中科院分区：
文献类型：
--
作者：
Hong, Vu;Presolski, Stanislav I.;Ma, Celia;Finn, M. G.
自 2002 年被发现以来，铜催化的叠氮化物-炔环加成 (CuAAC)[1] 反应（最广泛认可的点击化学示例 [2]）已迅速在众多领域得到应用。[3]该过程（及其无铜应变炔烃变体[4]）的吸引力源于叠氮化物和炔烃仅彼此之间的选择性反应性。由于生物分子的脆弱性和经常操作的低浓度，生物共轭对任何连接方法都提出了重大挑战。据报道，几种不同的 CuAAC 程序可用于解决涉及肽、蛋白质、多核苷酸和固定细胞的特定情况，通常会取得优异的结果，[5]，但偶尔也会得到不太令人满意的结果。 [6]我们在这里描述了一个普遍适用的程序，它解决了我们实验室中最棘手的点击生物共轭问题，因此应该在许多其他情况下使用。CuAAC 反应需要铜催化剂，通常用适当的螯合配体制备，[7] 保持在 CuI 氧化态。几年前，我们开发了一种以磺化红菲绕啉配体为特色的系统，[8]该系统被优化为有用的生物共轭方案。[9]一个显着的缺点是催化剂对氧的敏感性很高，需要无空气技术，当没有惰性气氛手套箱或在少量水溶液中使用敏感生物分子时，这种技术可能很难执行。我们还引入了一种电化学方法来生成和保护具有催化活性的 CuI-配体物质，用于 CuAAC 生物共轭和合成偶联反应，并且只需极少的努力即可排除空气。 [10]在这些条件下，氧气洗涤过程中不会产生过氧化氢，从而产生未受氧化副产物污染的蛋白质缀合物。然而，该解决方案也仅适用于能够使用适当设备的专家。其他方案采用铜 (I) 源，例如 CuBr 来标记固定细胞 [11] 和合成糖蛋白。 [12]在这些情况下，CuI 在空气中的不稳定性要求大
Since its discovery in 2002, the copper-catalyzed azide-alkyne cycloaddition (CuAAC)[1] reaction—the most widely recognized example of click chemistry [2]—has been rapidly embraced for applications in myriad fields.[3] The attractiveness of this procedure (and its copper-free strained-alkyne variant [4]) stems from the selective reactivity of azides and alkynes only with each other. Because of the fragile nature and low concentrations at which biomolecules are often manipulated, bioconjugation presents significant challenges for any ligation methodology. Several different CuAAC procedures have been reported to address specific cases involving peptides, proteins, polynucleotides, and fixed cells, often with excellent results,[5] but also occasionally with somewhat less satisfying outcomes.[6] We describe here a generally applicable procedure that solves the most vexing click bioconjugation problems in our laboratory, and therefore should be of use in many other situations.The CuAAC reaction requires the copper catalyst, usually prepared with an appropriate chelating ligand,[7] to be maintained in the CuI oxidation state. Several years ago we developed a system featuring a sulfonated bathophenanthroline ligand,[8] which was optimized into a useful bioconjugation protocol.[9] A significant drawback was the catalyst’s acute oxygen sensitivity, requiring air-free techniques which can be difficult to execute when an inert-atmosphere glove box is unavailable or when sensitive biomolecules are used in small volumes of aqueous solution. We also introduced an electrochemical method to generate and protect catalytically active CuI–ligand species for CuAAC bioconjugation and synthetic coupling reactions with miminal effort to exclude air.[10] Under these conditions, no hydrogen peroxide was produced in the oxygen-scrubbing process, resulting in protein conjugates that were uncontaminated with oxidative byproducts. However, this solution is also practical only for the specialist with access to the proper equipment. Other protocols have employed copper (I) sources such as CuBr for labeling fixed cells [11] and synthesizing glycoproteins.[12] In these cases, the instability of CuI in air imposes a requirement for large
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影响因子：
3.2
作者：
Bruckman, Michael A.;Kaur, Gagandeep;Wang, Qian
通讯作者：
Wang, Qian
DOI：
10.1007/bf01242085
发表时间：
1992-01-01
期刊：
MIKROCHIMICA ACTA
影响因子：
--
作者：
BAKER, WL;GOODE, J;COOPER, L
通讯作者：
COOPER, L
影响因子：
15
作者：
Lewis, WG;Magallon, FG;Finn, MG
通讯作者：
Finn, MG
影响因子：
15
作者：
Lallana, Enrique;Fernandez-Megia, Eduardo;Riguera, Ricardo
通讯作者：
Riguera, Ricardo
DOI：
10.1073/pnas.0707090104
发表时间：
2007-10-23
影响因子：
11.1
作者：
Baskin, Jeremy M.;Prescher, Jennifer A.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.