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公开(公告)号：US20140220633A1
公开(公告)日：2014-08-07
申请号：US14246783
申请日：2014-04-07
发明人： Roberto Falkenstein , Birgit Weydanz , Nicole Fuehrler , Claudia Giessel , Sybille Gabel , Adelbert Grossmann , Friederike Hesse , Marc Pompiati , Andreas Schaubmar , Brigitte Kraemer
摘要： The current invention reports a method for the purification of a not-glycosylated, heterologous polypeptide, which has been recombinantly produced in a prokaryotic cell, wherein the method comprises three chromatography steps of which the first chromatography step selected from i) hydrophobic charge induction chromatography, or ii) hydrophobic interaction chromatography, or iii) affinity chromatography, or iv) ion exchange chromatography, the second chromatography step is selected from i) anion exchange chromatography, or ii) cation exchange chromatography, or iii) hydroxylapatite chromatography, or iv) hydrophobic interaction chromatography, and the a third chromatography step is selected from i) hydrophobic charge induction chromatography, or ii) anion exchange chromatography, or iii) cation exchange chromatography, or iv) hydrophobic interaction chromatography, whereby the first chromatography step is an affinity chromatography in case of polypeptides capable of interacting with metal ligands, the second chromatography step is not a hydroxylapatite chromatography step in case of polypeptides with an isoelectric point below 6.0, and the third chromatography step can be performed in flow-through mode with polypeptides having a low or high isoelectric point.
摘要翻译： 本发明报道了在原核细胞中重组产生非糖基化的异源多肽的纯化方法，其中该方法包括三个色谱步骤，其中第一色谱步骤选自i）疏水电荷诱导色谱法， 或ii）疏水相互作用层析，或iii）亲和色谱法，或iv）离子交换色谱法，第二层析步骤选自i）阴离子交换色谱法，或ii）阳离子交换色谱法，或iii）羟基磷灰石色谱法，或iv）疏水性 并且第三层析步骤选自i）疏水电荷诱导色谱法，或ii）阴离子交换色谱法，或iii）阳离子交换色谱法，或iv）疏水相互作用色谱法，其中第一层析步骤是亲和层析 能够与met相互作用的多肽的情况 第二层析步骤在等电点低于6.0的多肽的情况下不是羟基磷灰石层析步骤，并且第三层析步骤可以以具有低或高等电点的多肽的流通模式进行。
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公开(公告)号：US09481706B2
公开(公告)日：2016-11-01
申请号：US14246783
申请日：2014-04-07
发明人： Roberto Falkenstein , Birgit Weydanz , Nicole Fuehrler , Claudia Giessel , Sybille Gabel , Adelbert Grossmann , Friederike Hesse , Marc Pompiati , Andreas Schaubmar , Brigitte Kraemer
摘要： The current invention reports a method for the purification of a not-glycosylated, heterologous polypeptide, which has been recombinantly produced in a prokaryotic cell, wherein the method comprises three chromatography steps of which the first chromatography step selected from i) hydrophobic charge induction chromatography, or ii) hydrophobic interaction chromatography, or iii) affinity chromatography, or iv) ion exchange chromatography, the second chromatography step is selected from i) anion exchange chromatography, or ii) cation exchange chromatography, or iii) hydroxylapatite chromatography, or iv) hydrophobic interaction chromatography, and the a third chromatography step is selected from i) hydrophobic charge induction chromatography, or ii) anion exchange chromatography, or iii) cation exchange chromatography, or iv) hydrophobic interaction chromatography, whereby the first chromatography step is an affinity chromatography in case of polypeptides capable of interacting with metal ligands, the second chromatography step is not a hydroxylapatite chromatography step in case of polypeptides with an isoelectric point below 6.0, and the third chromatography step can be performed in flow-through mode with polypeptides having a low or high isoelectric point.
摘要翻译： 本发明报道了在原核细胞中重组产生非糖基化的异源多肽的纯化方法，其中该方法包括三个色谱步骤，其中第一色谱步骤选自i）疏水电荷诱导色谱法， 或ii）疏水相互作用层析，或iii）亲和色谱法，或iv）离子交换色谱法，第二层析步骤选自i）阴离子交换色谱法，或ii）阳离子交换色谱法，或iii）羟基磷灰石色谱法，或iv）疏水性 并且第三层析步骤选自i）疏水电荷诱导色谱法，或ii）阴离子交换色谱法，或iii）阳离子交换色谱法，或iv）疏水相互作用色谱法，其中第一层析步骤是亲和层析 能够与met相互作用的多肽的情况 第二层析步骤在等电点低于6.0的多肽的情况下不是羟基磷灰石层析步骤，并且第三层析步骤可以以具有低或高等电点的多肽的流通模式进行。
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