重组人γ-干扰素包涵体稀释复性  

RENATURATION OF RECOMBINANT HUMAN INTERFERON-γ INCLUSION BODY BY DILUTION

作　　者：靳挺[1] 关怡新[1] 费峥峥[1] 姚善泾[1] 

机构地区：[1]浙江大学化学工程与生物工程学系,浙江杭州310027

出　　处：《化工学报》2004年第5期770-774,共5页CIESC Journal

基　　金：国家自然科学基金 (No 2 0 0 760 42 );浙江省自然科学基金 (No 2 0 10 99)资助项目~~

摘　　要：采用稀释法研究了重组人γ 干扰素包涵体复性条件 ,如温度、pH值、蛋白浓度、复性液中脲浓度 ,同时对加样操作方式进行了考察 ,得到了较为适宜的复性条件 .结果表明 ,脲能有效地抑制复性过程中蛋白质的聚集 ,合适的脲浓度随复性蛋白浓度的增加而有所增大 ,在 4℃、pH值 8 0、脲浓度 1 0mol·L-1、蛋白浓度0 1mg·ml-1及脉冲加样操作方式等条件下 ,复性后重组人γ 干扰素的活性为 2 39× 10 5IU·ml-1,比活达 2 2 1× 10 7IU·mg-1.The experimental conditions, including temperature, pH, denatured protein concentration and urea concentration, in the renaturation process of recombinant human interferon-γ inclusion body by dilution were studied and at the same time the pulse renaturation was tested. The optimum renaturation conditions were obtained. Results showed that urea could suppress the aggregation of protein during the renaturation process and the suitable urea concentration increased with the increase of denatured protein concentration. The activity and specific bioactivity was up to 2.39×105 IU&middot ml-1 and 2.21×107 IU&middotmg-1 respectively under 4°C, pH 8.0, 1.0 mol&middotL-1 urea, 0.1 mg&middotml-1 protein and pulse renaturation operation mode.

关 键 词：重组人Γ-干扰素 包涵体 稀释复性 

分 类 号：Q511[生物学—生物化学]