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目的:建立一种以κ-卡拉胶寡糖为配体,Affi-gel15为载体的亲和探针,用于分离纯化κ-卡拉胶寡糖在细胞信号通路中的特异性受体。方法:将标准寡糖和κ-卡拉胶寡糖进行己二胺衍生化后与Affi-gel15结合,并利用MS、NMR、PAGE电泳等方法对各步的产物进行分析。结果:MS、NMR结果表明,标准寡糖和κ-卡拉胶寡糖均能与己二胺进行完全稳定的衍生化反应,其衍生物与Affi-gel15的结合率都能达到50%以上,其中κ-卡拉胶寡糖-己二胺与Affi-gel15的结合率达到了68%。结论:制备得到的以κ-卡拉胶寡糖为配体的亲和探针能够与κ-卡拉胶寡糖的受体结合,如碱性成纤维细胞生长因子(bFGF)。这表明这种制备方法是可行的。
OBJECTIVE: To establish a affinity probe with κ-carrageenan oligosaccharide as ligand and Affi-gel15 as carrier, and to purify κ-carrageenan oligosaccharide specific receptor in cell signaling pathway. Methods: The standard oligosaccharides and kappa-carrageenan oligosaccharides were derivatized with hexamethylenediamine and then combined with Affi-gel15. The products of each step were analyzed by MS, NMR and PAGE. Results: The results of MS and NMR showed that all of the standard oligosaccharides and κ-carrageenan oligosaccharides were completely derivatized with hexamethylenediamine, and the binding rate of their derivatives to Affi-gel15 could reach more than 50% The binding rate of κ-carrageenan-hexamethylenediamine to Affi-gel15 reached 68%. CONCLUSION: The prepared affinity probe with κ-carrageenan oligosaccharide as a ligand can bind to κ-carrageenan oligosaccharides such as basic fibroblast growth factor (bFGF). This shows that this method of preparation is feasible.