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学科主题: 分析化学
题名: Immobilized enzyme reactors and their application in proteome study
作者: Zhang LH(张丽华) ;  Zhang YK(张玉奎)
会议文集: 26th International Symposium on Microscale Bioseparations
会议名称: 26th International Symposium on Microscale Bioseparations
会议日期: 2011-5-1
出版日期: 2011
会议地点: 圣地亚哥
通讯作者: 张丽华
出版者: 待补充
出版地: 待补充
合作性质: 分会特邀报告
部门归属: 1810
主办者: CASSS
摘要: Proteomics, as a new subject to understand various biological problems, is directed toward the identification of all proteins in cells, tissues or body fluids. At present, the “bottom-up” approach is widely applied for proteome study, by which protein digestion is an indispensable step. However, the traditional solution-based protein digestion has several drawbacks, such as long digestion time, unavoidable enzyme autodigestion and off-line operation. To solve these problems, recently much attention has been paid to the development of immobilized enzyme reactors (IMERs), which not only have high digestion efficiency, but also can be easily coupled with separation and detection systems to achieve automatic and high-throughput protein analysis. In our recent work, much effort has been made to design IMERs with various enzymes immobilized on different matrices. For the matrices, polymer particles, including acrylic polymer microparticles with either amino or epoxy groups, organic-inorganic hybrid silica monoliths, polymer monoliths, such as poly (acrylamide-co-methylenebisacrylamide) and poly (N-acryloxysuccinimide-co-poly (ethylene glycol) diacrylate), were applied, among which the hydrophilic polymer monoliths demonstrated the superiority of low non-specific adsorption of proteins or peptides. For enzymes, trypsin, chymotrypsin, pepsin and PNGase F were respectively immobilized. For the enzyme immobilization, either covalent bonding or metal chelation was used. By the former means, the immobilized enzyme was stable, and could be repeatedly used over months. By the latter means, the immobilized enzyme could be easily regenerated, to ensure the well-kept enzymatic activity of IMERs. All the above mentioned IMERs were successfully applied for the digestion of proteins, membrane proteins or glycoproteins, by which the protein digestion time could be obviously shortened from over 10 h to a few minutes or seconds. Furthermore, such IMERs were on-line integrated with protein separation and peptide separation by HPLC, to ensure the high throughput analysis of proteomes.
英文摘要: Proteomics, as a new subject to understand various biological problems, is directed toward the identification of all proteins in cells, tissues or body fluids. At present, the “bottom-up” approach is widely applied for proteome study, by which protein digestion is an indispensable step. However, the traditional solution-based protein digestion has several drawbacks, such as long digestion time, unavoidable enzyme autodigestion and off-line operation. To solve these problems, recently much attention has been paid to the development of immobilized enzyme reactors (IMERs), which not only have high digestion efficiency, but also can be easily coupled with separation and detection systems to achieve automatic and high-throughput protein analysis. In our recent work, much effort has been made to design IMERs with various enzymes immobilized on different matrices. For the matrices, polymer particles, including acrylic polymer microparticles with either amino or epoxy groups, organic-inorganic hybrid silica monoliths, polymer monoliths, such as poly (acrylamide-co-methylenebisacrylamide) and poly (N-acryloxysuccinimide-co-poly (ethylene glycol) diacrylate), were applied, among which the hydrophilic polymer monoliths demonstrated the superiority of low non-specific adsorption of proteins or peptides. For enzymes, trypsin, chymotrypsin, pepsin and PNGase F were respectively immobilized. For the enzyme immobilization, either covalent bonding or metal chelation was used. By the former means, the immobilized enzyme was stable, and could be repeatedly used over months. By the latter means, the immobilized enzyme could be easily regenerated, to ensure the well-kept enzymatic activity of IMERs. All the above mentioned IMERs were successfully applied for the digestion of proteins, membrane proteins or glycoproteins, by which the protein digestion time could be obviously shortened from over 10 h to a few minutes or seconds. Furthermore, such IMERs were on-line integrated with protein separation and peptide separation by HPLC, to ensure the high throughput analysis of proteomes.
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/116021
Appears in Collections:中国科学院大连化学物理研究所_会议论文

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Recommended Citation:
Zhang LH,Zhang YK. Immobilized enzyme reactors and their application in proteome study[C]. 见:26th International Symposium on Microscale Bioseparations. 圣地亚哥. 2011-5-1.
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