高等学校化学学报

高等学校化学学报 ›› 2004, Vol. 25 ›› Issue (7): 1257.

• 研究简报 • 上一篇    下一篇

纳米增强型毛细管酶柱用于葡萄糖液滴生物传感器的研究

冯锋1,2, 王柯敏1, 谌其亭1, 向秋芬1, 陈泽忠1,2, 黄红梅1, 黄杉生1, 霍希琴1   

  1. 1. 湖南大学生物传感与化学计量学国家重点实验室, 长沙 410082;
    2. 山西雁北师范学院化学系, 大同 037000
  • 收稿日期:2003-05-05 出版日期:2004-07-24 发布日期:2004-07-24
  • 通讯作者: 王柯敏(1957年出生),男,博士,教授,博士生导师,从事生物传感器研究.E-mail:kmwang@hnu.cn E-mail:kmwang@hnu.cn
  • 基金资助:

    国家自然科学基金重点项目(批准号:20135010);国家杰出青年基金(批准号:29825110);山西省自然科学基金(批准号:20011004);山西省重点建设学科资助

Application of Capillary Enzyme Column Enhancd by Using Nanoparticles for Glucose Biosensor Based on Liquid Droplets

FENG Feng1,2, WANG Ke-Min1, CHEN Qi-Ting1, XIANG Qiu-Fen1, CHEN Ze-Zhong1,2, HUANG Hong-Mei1, HUANG Shan-Sheng1, HUO Xi-Qin1   

  1. 1. State Key Lab of Chem-Biological Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China;
    2. Department of Chemistry, Shanxi Yanbei Normal Institute, Datong 037000, China
  • Received:2003-05-05 Online:2004-07-24 Published:2004-07-24

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被引次数

27

摘要： 葡萄糖的检测在临床医学以及食品工业等领域中十分重要.以往的检测方法主要包括化学发光法[1]、吸光光度法[2]、电化学法[3]和荧光法[4]等.固定化酶柱的制作是发展葡萄糖传感器的关键技术之一.传统的固定化方法主要是将具有生物活性的酶通过物理吸附、共价键合和交联的方法固定于载体基质上或包埋于有机聚合物的基质中.近期研究[5,6]表明,采用溶胶凝胶(Sol-gel)法将蛋白质和酶等生物活性物质包埋于无机陶瓷或玻璃材料内,保持生物组分的活性,且SiO2作为基质材料具有较好的坚固性、抗磨性、化学惰性以及高的光稳定性和透过性,但目前该法多用于电化学型生物传感器[7,8].本文利用纳米颗粒的比表面积大和吸附能力强等特点,将酶吸附在SiO2纳米颗粒表面,用易成膜的聚乙烯醇缩丁醛(PVB)作辅助基质在毛细管上固定酶,并采用分立式酶柱,克服了以往混合型酶柱普遍存在的酶促效率不高和使用寿命较短的局限性.所制得的酶柱具有表面反应活性高、表面活性中心多和催化效率高等特点.结合自行设计的液滴光化学传感装置[9,10],建立了一种高效、快速、微量的葡萄糖实时检测方法.

关键词: 纳米颗粒, 毛细管酶柱, 液滴传感器, 荧光分析法

Abstract: An effective and fast method for assay of micro-amounts of glucose was set up. A new technique for preparation of enzyme columns based on enzyme immobilization by sol-gel was investigated. Glucose oxidase(GOD) and horseradish peroxidase(HRP) mixed with SiO2 nanoparticles and polyvinyl butyral(PVB) medium were immobilized on the surface of capillary tube, respectively. The experimental results show that nanoparticles can significantly enhance the catalytic activity of immobilized enzyme. Based on GOD column and HRP column, a liquid droplet sensor was developed for the determination of glucose solution. The effect of separated columns and mixed columns on the response of glucose sensor was investigated. The sensor showed a linear response in a range of 2-400 ng/mL with a detection limit of 0.3 ng/mL under the optimum conditions. The characteristics of the sensor including effect of flow-rate, pH and temperature were discussed.

Key words: Nanoparticle, Capillary enzyme column, Liquid drop sensor, Fluorometry

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