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学科主题: 分析化学
题名: Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis
其他题名: 基于二磷酸腺苷(ADP)检测的电泳介导微分析技术用于己糖激酶抑制剂的筛选
作者: Wang TD(汪彤丹) ; Kang JW(康经武)
通讯作者: 康经武
刊名: Electrophoresis
发表日期: 2009
卷: 30, 期:8, 页:1349-1354
收录类别: SCI
部门归属: 上海有机所; 中科院研究生院
英文摘要: A CE-based method for hexokinase inhibitor screening was developed in the present paper, in this method, hexokinase activity was assayed via electrophoretically mediated microanalysis (EMMA), which combines on-column hexokinase-mediated reaction and measurement of produced adenosine 5'-diphosphate (ADP) via electrophoretical separation and UV detection. Enzyme inhibition can be read out directly from the reduced peak area of ADP in comparison with a reference electropherogram obtained in the absence of any inhibitor. Conditions for on-column enzyme reaction and separation of adenosine 5'-triphosphate (ATP) and ADP were optimized. The optimal buffer composition for enzymatic reaction was 25 mM HEPFS buffer (pH 7.5) containing 5 mM MgCl2, whereas the optimal buffer composition for separation was 100 in M Tris-phosphate buffer (pH 5.5) containing 0.02% (m/v) hexadimethrine bromide (HDB). Fortunately, discontinuous buffer system call be adapted easily in the EMMA method. The time for separation was reduced dramatically to less than 3 min by reversing the direction of EOF via dynamically coating the capillary wall with the cationic polyelectrolyte HDB. Moreover, the peak tailing of ATP was also reduced by HDB coating. The Z' factor as high as 0.98 was obtained, indicating a high quality of the screening data. The present method is simple, robust and cost-effective.
语种: 英语
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WOS记录号: WOS:000265968300009
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内容类型: 期刊论文
URI标识: http://ir.sioc.ac.cn/handle/331003/28685
Appears in Collections:分析化学研究室_期刊论文

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Recommended Citation:
Wang TD,Kang JW. Hexokinase inhibitor screening based on adenosine 5 '-diphosphate determination by electrophoretically mediated microanalysis[J]. Electrophoresis,2009,30(8):1349-1354.
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