摘要: |
采用超声破碎的方法提取hIFN-γ包涵体。经盐酸胍溶解、稀释复性、浓缩后通过SP Sepharose F.F层析纯化方法,简化了纯化步骤,缩短周期,提高了蛋白回收率。SDS-PAGE 检测表明,hIFN-γ纯度达97 %, 比活性为3.5?107 IU/mg,总回收率达40 %。说明整个工艺适合于大规模生产的要求,具有实际应用价值,从而为hIFN-γ批量生产奠定了良好的基础。 |
关键词: 重组人γ-干扰素,包涵体,复性,纯化 |
DOI: |
投稿时间:2011-10-04修订日期:2011-10-31 |
基金项目:国家自然科学资助(30972188) |
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Study on the Purification Technology of Recombinant Human Interferon-γ |
XU Chong-Li |
(College of Environment and Bioengineering,Jilin Institute of Chemical Technology,Jilin) |
Abstract: |
Inclusion body of hIFN-γ was obtained by the ultrasonication, renatured and concentrated by using guanidine hydrochloride denaturation. After the purification of SP Sepharose F.F, the total protein recovery was increased by simplifying the procedures and shortening the cycle. The product showed high purity exceeding 97% and the total protein recovery rate was nealy 40% by SDS-PAGE detecting. The high specific activity was 3.5?107 IU/mg. The production process reported here is so simple and convenient that it can be applied in the large scale production of INF-γ. This purification technology established a good foundation for the hIFN-γ mass production. |
Key words: Recombinant human Interferon-γ,inclusion body,renaturation,purification |