摘要
目的制备表面功能化磁性微球用以分离血管内皮生长因子(VEGF)核酸和固定化乳糖酶。方法采用悬浮聚合法分别进行含纳米氧化铁粒子的丙烯酸甲酯-苯乙烯和甲基丙烯酸环氧丙酯-苯乙烯的共聚,得到表面羧基化和表面环氧基化磁性微球;采用种子微乳液法制备氯甲基化磁性微球。用放射免疫测定方法研究磁性微球与b-γIgG的交联熏并对羧基化磁性微球进行VEGF核酸的磁性分离与乳糖酶的固定化实验。结果透射电子显微镜(TEM)、能量色散谱(EDS)和红外(IR)的结果显示制得的高分子磁性微球呈单分散状态,磁性纳米粒子在微球中分布均匀,表面含有特征性功能基团。放射免疫测定显示不同基团的磁性微球均能与b-γIgG交联,其中羧基化磁性微球达到75μg/mg的吸附量。羧基化磁性微球偶联相应的引物链可用于分离VEGF核酸探针。此外,固定化乳糖酶实验结果显示固定化酶的量与磁性微球的量成正比。结论表面功能化高分子磁性微球能与生物活性物质结合,在生物学和医学领域有广泛的应用前景。
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 - γIgG were determined by radioimmunoassay(RIA),and the separation of VEGF nucleic acid and lactase enzyme immobilizition 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 - γIgG and the absorption of b - γIgG reached75μg / mg,especially for the carboxyl - modi - fied magnetic microspheres.The carboxyl - modified magnetic microspheres can be used for the separ - ation 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 sur - face functional magnetic polymer microspheres can be bound with active bio - substance,and have a wide application prospect in the fields of biology and medicine.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2002年第2期118-123,共6页
Acta Academiae Medicinae Sinicae
基金
上海市新材料中心重点项目(99JC14016)资助
关键词
磁性微球
分离
核酸
固定化酶
magnetic microspheres
separation
nucleic acid
enzyme immobilization
