高等学校化学学报 ›› 2012, Vol. 33 ›› Issue (07): 1394-1400.doi: 10.3969/j.issn.0251-0790.2012.07.006

• 研究论文: 无机化学 • 上一篇    下一篇

生物功能化纳米SiO2 微球的构建及对谷胱甘肽S-转移酶的捕获分离

邹雪艳1,3, 褚留杰2, 董烁2, 赵彦保1, 李宾杰2   

  1. 1. 河南大学特种功能材料重点实验室, 开封 475004;
    2. 河南大学细胞与分子免疫学实验室, 开封 475004;
    3. 河南大学基础实验教学中心, 开封 475004
  • 收稿日期:2011-09-19 出版日期:2012-07-10 发布日期:2012-07-10
  • 通讯作者: 赵彦保, 男, 博士, 教授, 主要从事生物纳米材料制备研究. E-mail: yanbaozhao@126.com
    李宾杰, 男, 博士, 副教授, 主要从事生物纳米材料研究. E-mail: lbj821@yahoo.com.cn E-mail:lbj821@yahoo.com.cn
  • 基金资助:

    国家自然科学基金(批准号: 20671029), 省部共建基金(批准号: SBGJ090515 )和国家"八六三"计划项目(批准号: 2009AA03Z326)资助.

Construction of Bio-functional Nanosized Silica Microspheres for Capture and Separation of Glutathione S-Transferase

ZOU Xue-Yan1,3, CHU Liu-Jie2, DONG Shuo2, ZHAO Yan-Bao1, LI Bin-Jie2   

  1. 1. Key Laboratory for Special Functional Materials, Henan University, Kaifeng 475004, China;
    2. Key Laboratory of Cellular and Molecular Immunology, Henan University, Kaifeng 475004, China;
    3. Basic Experiment Teaching Center, Henan University, Kaifeng 475004, China
  • Received:2011-09-19 Online:2012-07-10 Published:2012-07-10

摘要: 采用巯基丙基三甲氧基硅烷(MPS)一步水解法制备了表面带有巯基(-SH)的纳米SiO2微球(nSiO2-SH), 探讨了水/醇体积比、 反应温度、 MPS初始浓度及反应时间对nSiO2-SH微球形貌的影响, 并分析了反应机理. 制备的nSiO2-SH微球进一步与还原型谷胱甘肽(GSH)中的-SH反应生成双硫键(-S-S-), 在微球表面键合上GSH分子, 得到了生物功能化的纳米nSiO2-GSH微球. 通过扫描电子显微(SEM)、 透射电子显微镜(TEM)、 傅里叶变换红外光谱(FTIR)和热重分析仪(TG)对样品的表面形貌、 尺寸和组成等进行了表征. 利用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)检测样品对谷胱甘肽S-转移酶(GST)的分离效果, 结果表明, nSiO2-GSH微球能从混合蛋白中特异性吸附GST, 达到了分离GST的目的.

关键词: 巯基纳米SiO2, 生物功能化, 特异性吸附, 谷胱甘肽S-转移酶分离

Abstract: Nanosized silica microspheres with sulfydryl group(nSiO2-SH) were synthesized with(3-mercaptopropyl)trimethoxysilane(MPS) via one-step hydrolyzing process, and the effects of volume ratio of water to ethanol, reaction temperature, reaction time and the volume of MPS on the product morphologies were discussed. Additionally, the possible reaction mechanism was also proposed. The prepared nSiO2-SH microspheres reacted further with glutathione through disulfide bond(-S-S-) to produce nSiO2-GSH microspheres. Their morphology, size and ingredient were characterized by scanning electron microscopy(SEM), transmission electron microscopy(TEM), Fourier transform infrared spectrometry(FTIR) and thermogravimetric(TG) analysis. The behavior of capture and separation of glutathione S-transferase(GST) was detected by sodium dodecylsulfate-polyacrylamide gel electrophoresis(SDS-PAGE) method. The results show that GST can be specifically adsorbed on nSiO2-GSH microspheres in the mixed protein solution. Therefore, nSiO2-GSH microspheres have potential applications in the separation, purification and detection of the GST-tagged fusion proteins.

Key words: Silica with sulfydryl group, Bio-functionalization, Specific adsorption, Separation of glutathione S-transferase

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