发明申请
- 专利标题: PROCESS FOR PURIFICATION OF RECOMBINANT HUMAN GRANULOCYTE COLONY STIMULATING FACTOR
- 专利标题(中): 重组人粒细胞集落刺激因子的纯化方法
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申请号: US13378973申请日: 2010-06-07
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公开(公告)号: US20120093765A1公开(公告)日: 2012-04-19
- 发明人: Sandeep Somani , Sriram Padmanabhan
- 申请人: Sandeep Somani , Sriram Padmanabhan
- 申请人地址: IN Mumbai, Maharashtra
- 专利权人: LUPIN LIMITED
- 当前专利权人: LUPIN LIMITED
- 当前专利权人地址: IN Mumbai, Maharashtra
- 优先权: IN865/KOL/2009 20090616
- 国际申请: PCT/IN2010/000377 WO 20100607
- 主分类号: A61K38/19
- IPC分类号: A61K38/19 ; C07K1/36 ; C07K14/535 ; C07K1/14
摘要:
The present invention describes a novel process for large-scale purification of therapeutic grade quality of recombinant human GCSF from microbial cells, wherein the protein is expressed as inclusion bodies. The Inclusion bodies are solubilized and refolded under redox condition. The Redox condition is provided by using ascorbic acid, dehydroascorbic acid and reduced gluthathione. The process involves the novel use of aqueous two phase extraction step to purify refolded GCSF after removal of denaturant. After this step GCSF is further purified using chromatography techniques for removal of related impurities. The GCSF obtained has good purity and yields which are essential for a production scale process. The host cell related contaminants like proteins, DNA and endotoxins are reduced using the purification processes of the invention.
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