视黄醇结合蛋白的分离纯化  

Purification of retinol-binding protein

作　　者：王世春[1] 刘云[1] 李永辉[1] 徐琪寿[1] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850

出　　处：《生物技术通讯》2004年第1期32-35,共4页Letters in Biotechnology

基　　金：国家自然科学基金(30170801)

摘　　要：押在变性条件下,应用Sephacryl-100凝胶过滤和Source-30Q阴离子交换两步分离,实现了分离纯化性质不稳定、易于降解的视黄醇结合蛋白(RBP)之目的。最后经过分步缓慢复性,获得具有生物活性的RBP,为其单克隆抗体制备及最终应用于临床营养评价和相关疾病的诊断创造了条件。Because of the difficulties of isolation and purification of retinol-binding protein(RBP)from human plasma and urine of kidney patients,to obtain RBP from Escherichia coli by constructing engineering strain which can expresse RBP.After the expression and washing of RBP inclusion in urine buffer,RBP was purified by the combination of Sephacryl-100gel filtration and Source Q anion-exchange chromatography.Purified RBP refolding was performed in the presence of glu-tathione oxide shuffling system.Fluorescence tiltering assay of retinol binding showed that refolded RBP restored its bind-ing ability with retinol.

关 键 词：视黄醇结合蛋白 分离 纯化 复性 单克隆抗体 

分 类 号：Q51[生物学—生物化学] Q26