多层开管毛细管酶反应器的制备及在蛋白质酶切中的应用  

Preparation of Multi-layer Open Capillary Immobilized Enzyme Reactor and Its Application in Digestion of Proteins

作　　者：胡佳薇[1] 申烨华[1] 秦伟捷[2] 张养军[2] 钱小红[2] 

机构地区：[1]西北大学合成与天然功能分子化学教育部重点实验室,新药研究所,化学与材料科学学院,西安710069 [2]蛋白质组学国家重点实验室,北京蛋白质组研究中心,北京放射与辐射医学研究所,北京102206

出　　处：《高等学校化学学报》2012年第2期268-275,共8页Chemical Journal of Chinese Universities

基　　金：国家“九七三”计划项目(批准号:2010CB912701,2012CB910603);国家自然科学基金(批准号:30621063,20735005,20875101,20905077,20975059)资助

摘　　要：以石英毛细管作为酶固定化的载体,在毛细管内壁上逐步合成树枝形大分子聚酰胺-胺(PAMAM),再通过交联剂戊二醛将胰蛋白酶直接键合到该大分子的末端氨基上,并对酶固定化条件进行了优化,制备了多层酶反应器.利用该酶反应器对马心细胞色素C等蛋白质进行了酶切,并对酶切的条件进行了优化.实验结果表明,该固定化酶反应器具有较高的酶切效率、良好的重现性和稳定性,可用于蛋白质组学的研究.With the capillary as the carrier of enzyme immobilization,poly（amidoamine） was gradually bon-ded to the inner wall of the capillary,then the trypsin was attached at the amino group of poly（amidoamine） with glutaraldehyde,and finally the multi-layer open capillary immobilized enzyme reactor was prepared after enzyme immobilization conditions had been optimized.Cyt-c was chosen as a model protein for evaluation of the performance of the multi-layer open capillary immobilized enzyme reactor,and its digestion conditions were optimized.The results demonstrate that the multi-layer immobilized enzyme reactor has high efficiency,good reproducibility and stability,and can be further applied in future proteomics study.

关 键 词：开管柱 固定酶反应器 蛋白质酶切 树枝形大分子 

分 类 号：O657[理学—分析化学]