An affinity-based method for the purification of fluorescently-labeled biomolecules.
An affinity-based method for the purification of fluorescently-labeled biomolecules.
复制标题
一种基于亲和力的荧光标记生物分子纯化方法。
DOI：
10.1021/bc060130i
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发表时间：
2006
影响因子：
4.7
通讯作者：
Francis,MatthewB
中科院分区：
文献类型：
--
作者：
Nguyen,Trung;Joshi,NeelS;Francis,MatthewB
由于难以分离标记和未标记的生物分子混合物，一种通用的新方法被开发出来用于修饰蛋白的亲和纯化。采用含有β-环糊精基团的sepharose固体载体选择性地捕获修饰过的发色团蛋白，而未修饰的蛋白则保留在溶液中。树脂分离后，用竞争性的环糊精粘合剂（如金刚烷羧酸）处理样品，释放修饰后的蛋白质。该方法在多种染料中得到了证明，这些染料具有广泛的光谱特征和不同的化学结构。初步研究表明，该方法也可用于富集存在于蛋白水解消化中的修饰肽片段。这项技术有望加速新的蛋白质修饰反应的发展，并为蛋白质组学的应用提供有用的工具。
Due to the difficulty of separating mixtures of labeled and unlabeled biomolecules, a general new method for the affinity purification of modified proteins has been developed. A Sepharose-based solid support bearing β-cyclodextrin groups was used to capture chromophore-modified proteins selectively, while unmodified proteins remained in solution. After isolation of the resin, the modified proteins were released by treating the sample with a competitive cyclodextrin binder, such as adamantane carboxylic acid. This procedure was demonstrated for several dyes displaying a wide range of spectral characteristics and diverse chemical structures. Preliminary studies have shown that this method can also be used to enrich modified peptide fragments present in proteolytic digests. This technique is anticipated to accelerate the development of new protein modification reactions and could provide a useful tool for proteomics applications.