亲和色谱-氢化物发生原子荧光分光光度法纯化检测人血浆中的硒蛋白-P  

Purification and Detection of Selenoprotein-P in Human Plasma by Affinity Chromatography-HG-AFS

作　　者：包建民[1] 于晓燕[1] 刘微[1] 李优鑫[1] 

机构地区：[1]天津大学药物科学与技术学院天津市现代药物传递及功能高效化重点实验室,天津300072

出　　处：《分析测试学报》2012年第7期804-809,共6页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(21075088);天津大学自主创新基金资助项目(1102213)

摘　　要：开发了两步亲和色谱法:肝素-琼脂糖凝胶、Ni-琼脂糖凝胶色谱纯化人血浆中硒蛋白-P的方法,并采用氢化物发生-原子荧光分光光度法(HG-AFS)检测,成功搭建了硒蛋白-P的纯化检测平台。确定了亲和色谱纯化的最佳梯度洗脱条件,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)定性检测,得到了一定纯度的硒蛋白-P,其回收率达43.2%。HG-AFS方法的线性相关系数为0.999 1,检出限为0.09μg/L,日内精密度(RSD)为0.12%,日间精密度(RSD)为0.27%,加标回收率为95%～104%。该亲和色谱纯化方法简单易控、回收率高,HG-AFS检测灵敏度高,结果准确可靠。A method was developed for the purification and detection of selenoprotein - P in human plasma. The sample was gradient eluted by two successive steps of affinity chromatography, i.e. , Heparin - Sepharose and Ni - Sepharose chromatography. The quality of the purified selenoprotein - P was detected by HG - AFS. The optimum gradient elution conditions were determined and the sele- noprotein - P with a certain purity was obtained by SDS - PAGE. The recovery of the two steps of af- finity chromatography reached up to 43.2%. The correlation coefficient of HG - AFS was 0. 999 1, and the detection limit was 0. 09 μg/L. The RSDs were 0. 12% for within-day and 0.27% for inter- day. The recoveries were in the range of 95% - 104% . The proposed method was simple and sen- sitive, and the results .obtained were accurate.

关 键 词：硒蛋白-P 人血浆 肝素-琼脂糖凝胶 Ni-琼脂糖凝胶 亲和色谱 氢化物发生原子荧光分光光度法 

分 类 号：O657.3[理学—分析化学] O613.52[理学—化学]