发明授权
- 专利标题: Purification of a polypeptide compound having a polysaccharide binding domain by affinity phase separation
- 专利标题(中): 通过亲和相分离纯化具有多糖结合结构域的多肽化合物
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申请号: US08505860申请日: 1995-07-24
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公开(公告)号: US06174700B1公开(公告)日: 2001-01-16
- 发明人: Charles A. Haynes , Peter Tomme , Douglas G. Kilburn
- 申请人: Charles A. Haynes , Peter Tomme , Douglas G. Kilburn
- 主分类号: C12P2106
- IPC分类号: C12P2106
摘要:
A compound having a polysaccharide binding domain such as contained by a cellulose and essentially lacking in polysaccharidase activity is purified from other ingredients in a mixture using an affinity partition system. A mixture containing the compound is contacted with a system containing as a first phase an aqueous solution of oligosaccharide polymer such as cellulose and as a second phase a solution of a polymer such as a poly(ethylene glycol)-poly(propylene glycol) copolymer. The compound petitions into the first phase and binds to the oligosaccharide polymer, preferably with a Ka of 103 to 107, to form a complex. The complex is collected, and the compound is dissociated from the oligosaccharide polymer. The compound may be formed of a non-peptide chemical moiety or a peptide moiety linked to a polypeptide having the polysaccharide binding domain. The compound may also be a fusion polypeptide containing the polysaccharide binding domain linked through a protease recognition sequence to a macromolecule such as an enzyme, a hormone or an antibody. The macromolecule can be removed by using a protease to cleave the recognition sequence. Another partition system contains the oligosaccharide polymer and a phase separation inducing agent such as a sulfate or citrate salt that induces separation to produce different phases.
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