用大型反相高效液相制备色谱法分离基因工程白细胞介素－2

Purification of Recombinant Human Interleukin-2 by Large-Scale Preparative Reversed-Phase High Performance Liquid Chromatography

探索了大规模生产制备基因工程白细胞介素－２（ｒＩＬ－２）的新工艺，采用等度和线性梯度相结合的方法，用大型反相高效液相制备色谱法分离提纯ｒＩＬ－２，并用电泳仪等测量了提纯物的纯度，结果表明：用新方法制备的ｒＩＬ－２纯度大于９５％，冻干后成品比活大于１０^（１０）ＩＵ／ｇ，且上样量大，在实际生产中应用价值很高。

Interleukin-2 (IL-2), has now been purified from natural and recombinant sources by a variety of methods, including small scale preparative reversed-phase high performance liquid chromatography (RP-HPLC), but the yield is low, and these methods are not suited to prepare IL-2 in practice. In the present paper we describe a new method for purification of recombinant human IL-2 (rIL-2) by large-scale preparative RP-HPLC in detail. In this method rIL-2 has been purified by Sephacryl-200 and further purified by large-scale preparative RP-HPLC. Gradient elution was performed with acetonitrile and water containing 0. l% trifluoroacetic acid and without the addition of sodium dodecyl sulfate (SDS). The separation results showed three chromatographic peaks. SDS polyacrylamide-gel electrophoresis (SDS-PAGE) and HPLC analyses conclude that the third peak is rIL-2. Its purity is over 95%. Further analyses showed the specific activity of rIL-2 purified with this method is over 1010IU/g. The second peak also has some rIL-2 and can be purified for the second time.