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表面功能化磁性微球的制备及其在核酸分离和酶固定化中的应用

[Preparation of surface functional magnetic microspheres and their application in nucleic acid separation and enzyme immobilization].

作者信息

Zhu Yi-hua, Wang Qiang-bin, Gu Hong-chen, Wang Sheng-lin, Gao Feng

机构信息

Key Laboratory for Ultrafine Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237, China.

出版信息

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2002 Apr;24(2):118-23.

Abstract

OBJECTIVE

To Prepare surface functional magnetic microspheres for the separation of vascular endothelial growth factor (VEGF) nucleic acid and lactase enzyme immobilization.

METHODS

Using suspension polymerization methods to copolymerize MA-styrene containing magnetite nanoparticles and GMA-styrene also containing magnetite nanoparticles, respectively. Both the carboxyl-modified magnetic microspheres and epoxy-modified magnetic microspheres were obtained. In addition, the chloromethyl-modified magnetic microspheres were prepared by seedy microemulsion. The magnetic microspheres bound with b-gamma IgG were determined by radioimmunoassay (RIA), and the separation of VEGF nucleic acid and lactase enzyme immobilization were performed by carboxyl-modified magnetic microspheres.

RESULTS

Transmission electron microscopy (TEM), energy-dispersive spectroscopy (EDS) and infrared (IR) spectra showed that the products of polymer magnetic microspheres were monodispersed and that the magnetic particles were uniformly distributed in the microsphere with special functional group on the surface of the microsphere. RIA showed that three kinds of magnetic microspheres could be bound with b-gamma IgG and the absorption of b-gamma IgG reached 75 micrograms/mg, especially for the carboxyl-modified magnetic microspheres. The carboxyl-modified magnetic microspheres can be used for the separation of VEGF nucleic acid by coupling with corresponding primer. Moreover, the immobilized enzyme was proportional to the amount of the carboxyl-modified magnetic microspheres.

CONCLUSIONS

The surface functional magnetic polymer microspheres can be bound with active bio-substance, and have a wide application prospect in the fields of biology and medicine.

摘要

目的

制备用于分离血管内皮生长因子(VEGF)核酸和固定乳糖酶的表面功能化磁性微球。

方法

分别采用悬浮聚合法使含磁铁矿纳米颗粒的甲基丙烯酸甲酯-苯乙烯和含磁铁矿纳米颗粒的甲基丙烯酸缩水甘油酯-苯乙烯进行共聚,得到羧基修饰磁性微球和环氧基修饰磁性微球。此外,通过种子微乳液法制备氯甲基修饰磁性微球。采用放射免疫分析法(RIA)测定与β-γ免疫球蛋白结合的磁性微球,并用羧基修饰磁性微球进行VEGF核酸的分离及乳糖酶的固定化。

结果

透射电子显微镜(TEM)、能谱仪(EDS)和红外光谱(IR)表明,聚合物磁性微球产物呈单分散状态,磁性颗粒均匀分布于微球中,且微球表面带有特殊官能团。RIA显示,三种磁性微球均可与β-γ免疫球蛋白结合,β-γ免疫球蛋白的吸附量达75微克/毫克,羧基修饰磁性微球尤为明显。羧基修饰磁性微球通过与相应引物偶联可用于VEGF核酸的分离。此外,固定化酶的量与羧基修饰磁性微球的量成正比。

结论

表面功能化磁性聚合物微球可与活性生物物质结合,在生物医学领域具有广阔的应用前景。

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