Institutional Repository of Key Laboratory of Coastal Zone Environmental Processes, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences (KLCEP)
| Potentiometric determination of trypsin using a polymeric membrane polycation-sensitive electrode based on current-controlled reagent delivery | |
Chen, Yan1,2; Ding, Jiawang1; Qin, Wei1
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| 发表期刊 | BIOELECTROCHEMISTRY
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| ISSN | 1567-5394 |
| 2012-12-01 | |
| 卷号 | 88页码:144-147 |
| 关键词 | Potentiometry Polycation-sensitive Electrode Controlled Release Protease Trypsin |
| 产权排序 | [Chen, Yan; Ding, Jiawang; Qin, Wei] Chinese Acad Sci, Shandong Prov Key Lab Coastal Zone Environm Proc, YICCAS, Key Lab Coastal Zone Environm Proc,Yantai Inst Co, Yantai 264003, Shandong, Peoples R China; [Chen, Yan] Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China |
| 通讯作者 | Qin, W (reprint author), Chinese Acad Sci, Shandong Prov Key Lab Coastal Zone Environm Proc, YICCAS, Key Lab Coastal Zone Environm Proc,Yantai Inst Co, Yantai 264003, Shandong, Peoples R China. |
| 作者部门 | 中科院海岸带环境过程与生态修复重点实验室 |
| 英文摘要 | A potentiometric biosensor for the determination of trypsin is described based on current-controlled reagent delivery. A polymeric membrane protamine-sensitive electrode with dinonylnaphthalene sulfonate as cation exchanger is used for in situ generation of protamine. Diffusion of protamine across the polymeric membrane can be controlled precisely by applying an external current. The hydrolysis catalyzed with trypsin in sample solution decreases the concentration of free protamine released at the sample-membrane interface and facilitates the stripping of protamine out of the membrane surface via the ion-exchange process with sodium ions from the sample solution, thus decreasing the membrane potential, by which the protease can be sensed potentiometrically. The influences of anodic current amplitude, current pulse duration and protamine concentration in the inner filling solution on the membrane potential response have been studied. Under optimum conditions, the proposed protamine-sensitive electrode is useful for continuous and reversible detection of trypsin over the concentration range of 0.5-5 U mL(-1) with a detection limit of 0.3 U mL(-1). The proposed detection strategy provides a rapid and reagentless way for the detection of protease activities and offers great potential in the homogeneous immunoassays using proteases as labels. (C) 2012 Elsevier B.V. All rights reserved.; A potentiometric biosensor for the determination of trypsin is described based on current-controlled reagent delivery. A polymeric membrane protamine-sensitive electrode with dinonylnaphthalene sulfonate as cation exchanger is used for in situ generation of protamine. Diffusion of protamine across the polymeric membrane can be controlled precisely by applying an external current. The hydrolysis catalyzed with trypsin in sample solution decreases the concentration of free protamine released at the sample-membrane interface and facilitates the stripping of protamine out of the membrane surface via the ion-exchange process with sodium ions from the sample solution, thus decreasing the membrane potential, by which the protease can be sensed potentiometrically. The influences of anodic current amplitude, current pulse duration and protamine concentration in the inner filling solution on the membrane potential response have been studied. Under optimum conditions, the proposed protamine-sensitive electrode is useful for continuous and reversible detection of trypsin over the concentration range of 0.5-5 U mL(-1) with a detection limit of 0.3 U mL(-1). The proposed detection strategy provides a rapid and reagentless way for the detection of protease activities and offers great potential in the homogeneous immunoassays using proteases as labels. (C) 2012 Elsevier B.V. All rights reserved. |
| 文章类型 | Article |
| 资助机构 | National Natural Science Foundation of China [41176081]; Outstanding Youth Natural Science Foundation of Shandong Province [JQ200814]; Taishan Scholar Program of Shandong Province [TS20081159] |
| 收录类别 | SCI |
| 语种 | 英语 |
| 关键词[WOS] | BIOANALYTICAL APPLICATIONS ; ELECTROCHEMICAL ASSAY ; PROTEASE ACTIVITY ; POLYION SENSORS ; SUBSTRATE ; INHIBITORS ; BLOOD |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Life Sciences & Biomedicine - Other Topics ; Biophysics ; Electrochemistry |
| WOS记录号 | WOS:000311065000022 |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.yic.ac.cn/handle/133337/6036 |
| 专题 | 中国科学院海岸带环境过程与生态修复重点实验室 |
| 作者单位 | 1.Chinese Acad Sci, Shandong Prov Key Lab Coastal Zone Environm Proc, YICCAS, Key Lab Coastal Zone Environm Proc,Yantai Inst Co, Yantai 264003, Shandong, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Chen, Yan,Ding, Jiawang,Qin, Wei. Potentiometric determination of trypsin using a polymeric membrane polycation-sensitive electrode based on current-controlled reagent delivery[J]. BIOELECTROCHEMISTRY,2012,88:144-147. |
| APA | Chen, Yan,Ding, Jiawang,&Qin, Wei.(2012).Potentiometric determination of trypsin using a polymeric membrane polycation-sensitive electrode based on current-controlled reagent delivery.BIOELECTROCHEMISTRY,88,144-147. |
| MLA | Chen, Yan,et al."Potentiometric determination of trypsin using a polymeric membrane polycation-sensitive electrode based on current-controlled reagent delivery".BIOELECTROCHEMISTRY 88(2012):144-147. |
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