离子交换及明胶包被琼脂糖凝胶的制备与应用
文献类型：学位论文
| 作者 | 王国祥 |
| 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 |
| 导师 | 聂峰光 |
| 关键词 | 琼脂糖凝胶 离子交换介质 明胶 细胞培养微载体 |
| 其他题名 | Preparation and application of ion exchange and gelatin coated agarose gel |
| 学位专业 | 生化工程 |
| 中文摘要 | 该文利用琼脂糖良好的衍生性能，对交联琼脂糖凝胶提出了两种方法进行了修饰，制备了两种琼脂糖衍生介质.采用DEAE-HCl与琼脂糖上羟基进行非均相缩合反应，偶联上离子交换功能基团，制备了生物大分子纯化分离用弱阴离子型离子交换介质.考察了温度、时间、DEAE-HCl浓度以及NaOH浓度对偶联反应的影响.采用明胶包被修饰琼脂糖凝胶，经过交联固化后，制得了动物细胞培养微载体.用明胶包被琼脂糖微载体进行了Vero细胞培养实验，发现经过包被修饰后，琼脂糖凝胶具有良好的支持细胞贴壁生长性能，其培养状况与Cytodex-3相当.培养一周后，Vero细胞密度达到9.93×10<'5>cells/ml.载体肯有良好的颗粒性质、消化性能、回收性能，在培养过程中会出现载体间架桥现象，可以实现细胞在载体间的迁移传代. |
| 英文摘要 | Two agarose gel derivatives were prepared including a DEAE anion exchange medium for protein purification and a gelatin coated medium for cell culture. A weak anion exchange medium for protein purification was prepared by coupling the function group DEAE onto agarose gels. It was found that the reaction time, reaction temperature and the concentration of reactants such as DEAE-HC1, NaOH had great influence on the degree of coupling reaction. The optimum reaction conditions were 60℃ for 1 hour under 3mol/L DEAE-HC1, 3.5mol/L NaOH. The resulting capacity of the synthesized gel was 0.168mmol/ml gel, which was similar to that of the commercial product, DEAE Sepharose CL-6B. Evaluation was carried out on BSA adsorption equilibrium, acid-base titration curve and the dynamic binding capacity of the exchanger made in this study. The medium was used to purify a protein mixture of BSA and HB. Good purification was achieved. In the second approach, agarose gel was coated by gelatin to form a composite gel. This gel had the good property to support the cell growth on the surface when used as animal cell culture microcarrier. The gelatin-coated agarose had been used to culture Vero cells in M199 cell culture medium. After a week, the cell density on the microcarrier was up to 9.93 * 10~5cells/ml, which was comparable to the result of Cytodex-3. The recycling property of coated agarose was also investigated. It was found that treated by trypsin, the agarose matrix could be recycled and coated by gelatin again. The treated gel also had the good culture ability. During the culture, the 'bridging' effect among coated agarose enhanced the cells' ability to transfer from confluent beads to bare ones, which could be used to bead-to-bead cell transfer. The research about the preparation of agarose anion exchanger and gelatin-coated agarose was some novel, which was not reported. |
| 语种 | 中文 |
| 公开日期 | 2013-09-26 |
| 页码 | 70 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/1900]  |
| 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 | 王国祥. 离子交换及明胶包被琼脂糖凝胶的制备与应用[D]. 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
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