重组猪α-干扰素的纯化及其抗病毒活性测定

The purification and antiviral activity determination of recombinant porcine α-interferon(PoIFN-α)

分别采用PEG6000沉淀法和Sephadex G-50凝胶色谱法纯化毕赤酵母分泌表达的重组猪α-干扰素（PolFN-α）。正交试验结果显示：PEG6000沉淀法最佳分离条件为pH6．0、NaCl0．2mol／L、PEG60000．1g／mL，蛋白回收率约80％；纯化后蛋白效价为2．9×105IU／mL，比活为1．04x105IU／mg。通过SephadexG．50凝胶色谱纯化，重组PolFN-α的回收率为60％左右，纯度达到90％；纯化后蛋白效价为8×103IU／mL，比活为6．7×103IU／mg。

The recombinant porcine α-interferon （PoIFN-α）expressed by Pichia pastoris was purified by PEG 6000 precipitation and Sephadex G-50 gel chromatography respectively. The orthogonal experiment results showed that the optimal conditions of PEG 6000 precipitation was pH 6, 0.2 mol/L NaCI and 0.1 g/mL PEG 6000,in which the protein＇s recovery rate was about 80%, and after purified its valence and specific activity were 2.9 x l05 IU/mL and 1.04 x l0s IU/mg respectively. When the PoIFN-a was purified by Sephadex G-50 gel chromatography,its recovery rate and purity were about 60% and 90% respectively,and its valence and specific activity were 8 x 103 IU/mL and 6.7 x 103 IU/mg respectively.