| 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 |
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| 引用本文： | 胡佳薇,申烨华,秦伟捷,张养军,钱小红.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[J].高等学校化学学报,2012,33(2):268-275. |
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| 作者姓名： | 胡佳薇 申烨华 秦伟捷 张养军 钱小红 |
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| 作者单位： | 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 |
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| 基金项目： | 国家"九七三"计划项目，国家自然科学基金 |
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| 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.
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| 关 键 词： | 开管柱 固定酶反应器 蛋白质酶切 树枝形大分子 |
| 收稿时间： | 2011-02-07 |
Preparation of Multi-layer Open Capillary Immobilized Enzyme Reactor and Its Application in Digestion of Proteins |
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| HU Jia-Wei
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SHEN Ye-Hua
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QIN Wei-Jie
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ZHANG Yang-Jun
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QIAN Xiao-Hong.Preparation of Multi-layer Open Capillary Immobilized Enzyme Reactor and Its Application in Digestion of Proteins[J].Chemical Research In Chinese Universities,2012,33(2):268-275. |
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| Authors: | HU Jia-Wei SHEN Ye-Hua QIN Wei-Jie ZHANG Yang-Jun QIAN Xiao-Hong |
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| Institution: | 1. Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Drug Research Institute, College of Chemistry and Materials Science, Northwest University, Xi'an 710069, China;
2. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China |
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| Abstract: | 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. |
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| Keywords: | Open-tubular capillary Immobilized enzyme reactor Protein digestion Dendritical macromolecule |
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