高等学校化学学报

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微流控芯片上同工酶的孵育及活性检测

刘菁1,2; 庄贵生1,2; 贾春平1; 金庆辉1; 王惠民3; 赵建龙1,4; 杨梦苏1   

  1. 1. 中国科学院上海微系统与信息技术研究所, 上海 200050;
    2. 中国科学院研究生院, 北京 100039;
    3. 南通大学附属医院, 南通 226001;
    4. 生物芯片上海国家工程研究中心, 上海 201203
  • 收稿日期:2005-08-22 修回日期:1900-01-01 出版日期:2006-07-10 发布日期:2006-07-10
  • 通讯作者: 杨梦苏

Incubation and Activity Determination of Lactate Dehydrogenase Isoenzymes
on Microfluidic Chip

LIU Jing1,2; ZHUANG Gui-Sheng1,2; JIA Chun-Ping1; JIN Qing-Hui1;
WANG Hui-Min3; ZHAO Jian-Long1,4; YANG Meng-Su1*   

  1. 1. Shanghai Institute of Microsystem and Information Technology, Chinese
    Academy of Sciences, Shanghai 200050, China;
    2. Graduate School of Chinese Academy of Sciences, Beijing 100039, China;
    3. Affiliated Hospital of Nantong University, Nantong 226001, China;
    4. National Engineering Research Center for Shanghai Biochip Technology,
    Shanghai 201203, China
  • Received:2005-08-22 Revised:1900-01-01 Online:2006-07-10 Published:2006-07-10
  • Contact: YANG Meng-Su

摘要: 建立了一种在微流控芯片上进行同工酶孵育及活性检测的方法. 该方法在集成温控装置的微流控芯片上实现对同工酶与辅酶反应进程的控制, 完成同工酶的进样、孵育反应、电泳分离和活性检测的实验步骤. 建立了基于微流控芯片的同工酶荧光检测系统, 使用360 nm光源激发辅酶产生荧光, 在460 nm处选择性采集荧光信号. 在微流控芯片上实现了同工酶样品的快速活性检测, 酶活性检测限达到0.5 U/L.

关键词: 微流控芯片, 温控, 同工酶, 活性检测

Abstract: A novel incubation and activity determination method of lactate dehydrogenase isoenzymes was established. Based on the on\|chip electrophoretic separation theory the analyzer was integrated with a temperature\|controlling system. The assay program of the enzymes including injection of samples, incubation and separation was highly performed in the microchannel. A system for high sensitivity fluorescence detection was developed. A 360 nm light source was adopted to excite the fluorescence and the 460 nm signature was finally selectively gathered. The efficient separation of different LDH fractions indicates the potential of microfluidic devices for isoenzymes assay and the detection limit reached 0.5 U/L.

Key words: Microfluidic chip, Temperature-controlling, Lactate dehydrogenase isoenzymes, Activity determination

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