甲醇酵母分泌表达重组人干扰素α1b纯化工艺的建立

Establishment of purification procedure of recombinant human interferon-α1b secreted and expressed by engineering pichia pastoris

建立适合大规模、低成本生产重组人干扰素α1b(Recombinanthumaninterferon α1b ,rhIFN α1b)的纯化工艺。采用高效分泌表达rhIFN α1b的甲醇酵母工程菌发酵 ,收集离心后的上清液 ,超滤脱盐 ,经离子交换柱和分子筛柱层析纯化。纯化的rhIFN α1b的纯度为 98%以上 ,比活性 2 .4× 10 7IU/mg ,活性回收率 14 % ,相对分子质量 1980 0和等电点 5 .0。经检测 ,rhIFN α1b蛋白N 端 15个氨基酸序列与正常对照完全符合。该纯化工艺简便 ,时程短 ,重复性好 。

To establish a large-scale with low-cost purification procedure for producing recombinant human interferon-α1b (rhIFN-α1b),rhIFN-α1b expressed efficiently in supernatant by recombinant pichia pastoris was collected de-salted and purified by centrifugation,ultrafiltration,ion-exchange and gel filtrafion successively.The purity ,specific activity,recovery rate of activity,relative molecular weight (M.W)and PI were over 98%,2.4×10 7IU/mg, 14%,19800 and 5.0 respectively .The purification is simple ,timesaving,well reproducible and suitable for large-scale production of homan IFN-α1b.