发明名称 |
Methods for purification of recombinantly produced proteins |
摘要 |
Recovery of the 52/48 kDa tryptic fragment of vWF or peptide subfragments thereof produced in the form of inclusion bodies from recombinant host cells is carried out by providing a washed recombinant host cell suspension, adding a detergent and subjecting the cells to mechanical disruption. A second detergent is added, followed by another mechanical disruption and centrifugation to a pellet. The pellet is resuspended in a buffer and subjected to another mechanical disruption. Inclusion bodies are washed, resuspended and then recovered. In addition, endotoxins and DNA are removed from inclusion bodies containing the 52/48 kDa tryptic fragment of vWF or peptide subfragments thereof by mechanically disrupting the inclusion bodies in an aqueous buffer containing a detergent. A washed pellet is formed from these mechanically disrupted inclusion bodies and dissolved in a denaturant. Alkylation is then performed and the suspension is subjected to column chromatography, where the alkylated fragment or subfragment is recovered by elution.
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申请公布号 |
US5530100(A) |
申请公布日期 |
1996.06.25 |
申请号 |
US19900519709 |
申请日期 |
1990.05.07 |
申请人 |
RHONE-POULENC RORER PHARMACEUTICALS INC. |
发明人 |
DARLING, THOMAS L. J.;AKHNANA, LIDA Y.;MITSCHELEN, JONATHAN J.;HRINDA, MICHAEL E. |
分类号 |
C07K14/755;(IPC1-7):C07K3/08;C07K3/12;C07K3/20;C07K15/06 |
主分类号 |
C07K14/755 |
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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