【作者】 张岩；

【导师】 陈霞； 刘照惠；

【作者基本信息】 吉林大学 ， 生物工程， 2007， 硕士

【摘要】 白细胞介素-2是(Interleukin-2,IL-2)是Morgan等人于1976年首次发现的一种细胞因子,其主要成分是TH细胞释放的一种糖蛋白,分子量为15～17KD。随着现代生物技术的发展,它作为一种抗感染、抗肿瘤的基因工程药品,其特异稳定的临床疗效已得到社会认可。目前用于临床的IL-2是由工程菌(E.coli)所生产,而许多外原基因在原核细胞高效表达后,会形成不溶性的包涵体,IL-2也是这样。通过对包涵体进一步分离、纯化,使其具有与天然IL-2几乎相同的生物学活性。IL-2含有三个半胱氨酸,复性时由于二硫键错配可能会形成没有活性的异构体,这就使IL-2的复性在整个分离纯化过程中显得至关重要,它直接关系到产品的纯度和回收率。本研究将重点放在IL-2复性后,应用高效液相反相色谱技术去除IL-2错配异构体,以提高IL-2的生物学活性,同时对复性方法略加讨论。更多还原

【Abstract】 Interleukin2(IL-2) is an very important immune regulation molecule, it can increase the function of immune system. IL-2, as a gene engineer anti tumor drug, can be broadly used into clinical and the stable ,specific therapeutic effect are generally recognized by society. There are 3 cysteine residue for a natural conformation of IL - 2 and they can produce 3 kind of disulfide bond combination when oxidized, but only molecule that have disulfide bond between Cys-58 and Cys-105 can have biological activity. In the process of IL-2 purification, the process of renaturation is mostly important. It inevitably produce mismatch isomer and affect the purification and biological activity of IL-2. This research separate the 3 isomer of IL-2 with the technique of RP-HPLC and increased the specific activity and product efficiency and optimized IL-2 separation ,purification technology. This research pave the way for producing high level activity product and large scale commercial process. It also provide a theory and experiment foundation for further research.更多还原