基因工程白细胞介素2的复性条件和检测方法比较

从表达重组IL—2的E.coli中纯化IL—2,可因变性剂作用而使IL—2失去生物活性.本实验研究了IL—2浓度和复性剂浓度对复性的影响.结果表明,在IL—2浓度为0.05mg/mL、CuCl_2浓度为0.05mmol/L时,复性效果较好.本实验还比较了3种检测IL—2活性的方法,应用CTLL细胞检测,特异性高,但维持细胞费时费力;应用小鼠胸腺细胞和活化的小鼠脾细胞检测,简便易行,但敏感性远不如CTLL细胞检测.

Renatural Condition of rIL-2 and Comparision of IL-2 Bioas- say Methods

A study was undertaken to determine the effect of concentration of protein and renatural agent on IL-2 renaturation. The results showed that the renatural effect was excellent when the concentrations of protein and CuCl2 were 0. 05 mg/mL and 0.05 mmol/L, respectively. In the experiment, three methods of IL-2 bioassay have been compared. The method based on the proliferation of CTLL cells was quite specific but it was not easy to support the growth of the cell line. The proliferations of mouse thymus cells and activated mouse spleen cells were also used for IL-2 bioassay. These method were simple but their sensitivities were not so good as that of CTLL cells.