基因工程白介素2的纯化研究

Studies of purifying recombinant interleukin 2

由E.coli表达的重组IL-2(rIL-2)是以一种不溶性、呈聚合状态的包含体形式存在于E.coli的胞浆之中。在rIL-2的纯化研究中我们以包含体的制备为重点,经本实验所建立的包含体制备方法可使其rIL-2的纯度达86%,包含体溶解后的rIL-2纯度达90%,经Sephadex G200凝胶层析后的rIL-2纯度达98%。本实验还研究了蛋白浓度、pH、复性剂对rIL-2复性的影响。

Recombinant human interleukin-2 (rIL-2) Produced in E.coli was isolated as insoluble aggregates of protein (inclusion bodies IBs) after cell breakage. Before initiating disulfide bone formation in vitro, we isolated and purified the reduced rIL-2 by taking advantage of its insolubility. We used the methods to purify rIL-2 inclusion bodies that achieved 86% purity, after solubilization of IBs the purity was 90% and further purified in reduced and devatured form by gel-permention yielding 98% pure IL-2 witn biological activity recovery of 59%. Besides we had also studied the effects of protein concenfration, PH and re natural agent on refold and renaturation of reduced rIL-2.