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用于生物催化和生物分离的含磁铁矿球形二氧化硅纳米颗粒。

Magnetite-containing spherical silica nanoparticles for biocatalysis and bioseparations.

作者信息

Yang Huang-Hao, Zhang Shu-Qiong, Chen Xiao-Lan, Zhuang Zhi-Xia, Xu Jin-Gou, Wang Xiao-Ru

机构信息

Material Transport & Transformation in Environmental & Life Processes Lab, The First Institute of Oceanography, SOA, Qingdao, 266061, PR China.

出版信息

Anal Chem. 2004 Mar 1;76(5):1316-21. doi: 10.1021/ac034920m.

Abstract

The simultaneous entrapment of biological macromolecules and nanostructured silica-coated magnetite in sol-gel materials using a reverse-micelle technique leads to a bioactive, mechanically stable, nanometer-sized, and magnetically separable particles. These spherical particles have a typical diameter of 53 +/- 4 nm, a large surface area of 330 m(2)/g, an average pore diameter of 1.5 nm, a total pore volume of 1.427 cm(3)/g and a saturated magnetization (M(S)) of 3.2 emu/g. Peroxidase entrapped in these particles shows Michaelis-Mentan kinetics and high activity. The catalytic reaction will take place immediately after adding these particles to the reaction solution. These enzyme entrapping particles catalysts can be easily separated from the reaction mixture by simply using an external magnetic field. Experiments have proved that these catalysts have a long-term stability toward temperature and pH change, as compared to free enzyme molecules. To further prove the application of this novel magnetic biomaterial in analytical chemistry, a magnetic-separation immunoassay system was also developed for the quantitative determination of gentamicin. The calibration for gentamicin has a working range of 200-4000 ng/mL, with a detection limit of 160 ng/mL, which is close to that of the fluorescent polarization immunoassay (FPIA) using the same reactants.

摘要

利用反胶束技术将生物大分子与纳米结构的二氧化硅包覆磁铁矿同时包封于溶胶-凝胶材料中,可得到具有生物活性、机械稳定、纳米尺寸且可磁分离的颗粒。这些球形颗粒的典型直径为53±4 nm,比表面积为330 m²/g,平均孔径为1.5 nm,总孔体积为1.427 cm³/g,饱和磁化强度(Ms)为3.2 emu/g。包封于这些颗粒中的过氧化物酶表现出米氏动力学和高活性。将这些颗粒加入反应溶液后,催化反应立即发生。这些包埋酶颗粒催化剂可通过简单地使用外部磁场轻松地从反应混合物中分离出来。实验证明,与游离酶分子相比,这些催化剂对温度和pH变化具有长期稳定性。为了进一步证明这种新型磁性生物材料在分析化学中的应用,还开发了一种用于庆大霉素定量测定的磁分离免疫分析系统。庆大霉素的校准工作范围为200 - 4000 ng/mL,检测限为160 ng/mL,这与使用相同反应物的荧光偏振免疫分析(FPIA)相近。

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