超顺磁性高分子微球的修饰、表征及其亲和分离蛋白质的初步研究
文献类型：学位论文
| 作者 | 姜波 |
| 学位类别 | 硕士 |
| 答辩日期 | 2001-06 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 刘会洲 |
| 关键词 | 超顺磁性 高分子微球 聚乙二醇 聚醋酸乙烯酯 活性三嗪性染料 乙醇脱氢酶 亲和作用 |
| 其他题名 | Modification and Characterization of Superparamagnetic Polymer Microspheres and Its Application in Affinity Protein |
| 学位专业 | 生化分离科学与工程 |
| 中文摘要 | 该文在合成超顺磁性PVA高分子微球的基础上，对磁性高分子微球进行表面聚乙二醇改性，并对改性后的磁性高分子微球的主要性能进行了表征.其后，进一步引入具有亲和作用的功能性基因，研究了采用三嗪染料CB修饰后的亲和磁性高分子微球从较复杂的真实体系中分离纯化乙醇脱氢酶的可行性，包括酶和蛋白在载体上的吸附-解吸以及不同载体、不同pH、不同离子强度对吸附分离纯化乙醇脱氢酶的影响等. |
| 英文摘要 | In the present work, PEG-modified Fe_3O_4/PVA superparamagnetic polymer micropheres were synthesized and characterized by microphotography, Fourier Transfrom infrared spectroscopy(FT-IR) etc. Superparamagnetic PVA microspheres with and without PEG-modified then were surface modified with dye Cibacon Blue F3GA(CB). Their performance on affinity separation of yeast alcohol dehydrogenase(YADH) from yeast was systematically investigated. It was found that the PEG could be immobilized onto Fe_3O_4/PVA superparamagnetic polymer micropheres by polymer function reaction. The laser diffraction particle analysis indicates that the size of the micropheres has a Gaussian distribution. The magnetic polymer microspheres are not only stable in a wide range from pH2 to pH14, but also stable in a wide temperature range. The content of PEG and Fe_3O_4 were about 190mg/g and 93mg/g in the microspheres respectively so that the superparamagnetic polymer micropheres have good magnetism properity and new function. These properties are very useful in the field of biotechnology and catalysis. The affinity separation application on yeast alcohol dehydrogenase(YADH) from yeast with the Fe_3O_4/PVA superparamagnetic polymer micropheres was investigated before and after modification by PEG and CB. Adsorption amounts of protein and the activities of adsorbed YADH were measured under various adsorption carriers. The amount of protein adsorption was strongly affected by the modification of particles. The activities of adsorbed YADH on both PEG and CB modified particle were higher than those of on the CB modified only. PEG modification will decrease non-special adsorption on Fe_3O_4/PVA superparamagnetic polymer microphere, but it can improve process selectivity. With various carriers, the recovery of YADH activity obtained was ～50% and the purification factor were 3.6-fold by PEG-CB modified carrier after one cycle of adsorption and desorption. |
| 公开日期 | 2013-09-26 |
| 页码 | 77 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/1897]  |
| 专题 | 过程工程研究所_研究所（批量导入） |
| 推荐引用方式 GB/T 7714 | 姜波. 超顺磁性高分子微球的修饰、表征及其亲和分离蛋白质的初步研究[D]. 北京. 中国科学院研究生院. 2001. |
入库方式： OAI收割
来源：过程工程研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。