发明名称 |
Expression and purification of recombinant soluble tissue factor |
摘要 |
A method is disclosed to make any protein in a form that can be isolated rapidly from a solution using a specific monoclonal antibody designated "HPC-4". It has now been determined that it is possible to form a fusion protein of the epitope with a protein to be isolated, and isolate the protein using HPC-4-based affinity chromatography. In the preferred embodiment, a specific protease cleavage site is inserted between the epitope and the protein so that the epitope can be easily removed from the isolated protein. In an example, a functionally active soluble tissue factor including the twelve amino acid epitope recognized in combination with calcium by HPC-4 and a factor Xa cleavage site was expressed from a vector inserted into a procaryotic expression system. The recombinant tissue factor can be rapidly isolated in a single chromatographic step using the HPC-4 monoclonal antibody immobilized on a suitable substrate. Once isolated, the Protein C epitope is removed by cleavage with factor Xa, leaving the functionally active, soluble tissue factor.
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申请公布号 |
US5298599(A) |
申请公布日期 |
1994.03.29 |
申请号 |
US19920816679 |
申请日期 |
1992.01.03 |
申请人 |
OKLAHOMA MEDICAL RESEARCH FOUNDATION |
发明人 |
REZAIE, ALIREZA;ESMON, CHARLES T.;MORRISSEY, JAMES H. |
分类号 |
C12N15/09;A61K38/00;C07K1/107;C07K14/47;C07K14/745;C07K16/40;C07K19/00;C12N9/64;C12P21/02;C12R1/91;G01N33/68;G01N33/86;(IPC1-7):C07K13/00 |
主分类号 |
C12N15/09 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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