重组人干扰素α_(2b)和胸腺肽α_1融合蛋白的分离纯化  

Separation and purification of recombinant fusion proteins of IFNα_(2b)-THYα_1

作　　者：罗琪[1] 陈劲春[1] 

机构地区：[1]北京化工大学生命科学与技术学院,北京100029

出　　处：《北京化工大学学报（自然科学版）》2008年第5期82-84,共3页Journal of Beijing University of Chemical Technology(Natural Science Edition)

摘　　要：考察了大肠杆菌E.coli表达的重组人干扰素α2b和胸腺肽lα融合蛋白包涵体的变性、复性及融合蛋白的分离纯化过程。实验结果表明:包涵体经7 mol/L盐酸胍,10 mmol/L DTT变性;1 mol/L尿素,2 mmol/L还原型谷胱甘肽,0.2 mmol/L氧化型谷胱甘肽脉冲加样稀释复性;金属螯合层析收集0.3 mol/L咪唑洗脱峰,冷干后经重组肠激酶30℃酶切24 h、Sephadex G-50柱纯化,可得到纯度达90%的重组人干扰素α2b和胸腺肽αl融合蛋白,且最终目的融合蛋白产量达68 mg/g干菌体。An optimized method has been developed for separation and purification of recombinant fusion proteins of IFNα2b-THYα1 with transgenic E. coli. After denaturation and renaturation, the fusion proteins were purifled using three chromatographic columns--namely Ni-chelating-Sepharose before and after enzyme digestion, and Sephadex G-50-were operated in series. The purity of the resulting fusion proteins was more than 90 %, as shown by RP-HPLC analysis. The yield of recombinant fusion proteins can be as high as 68 mg/g dry thalli.

关 键 词：人干扰素α2b 胸腺肽αl 融合蛋白 金属螯合层析 

分 类 号：Q512.7[生物学—生物化学]