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1. 发明申请

WO2005019466A3 PROCESS FOR THE PREPARATION OF RECOMBINANT POLYPEPTIDES 审中-公开
公开(公告)号：WO2005019466A3
公开(公告)日：2005-03-03
申请号：PCT/EP2004/009321
申请日：2004-08-19
申请人： SANDOZ AG , STEMPFER, Günter , PALMA, Norbert
发明人： STEMPFER, Günter , PALMA, Norbert
IPC分类号： C12P21/02
摘要： The present invention relates to the preparation of a recombinant polypeptide, which polypeptide upon expression has been secreted into the periplasm of a transformed host cell. In particular, this invention relates to a process to enhance the extraction yield of said recombinant polypeptide from the periplasm before further downstream processing.

2. 发明申请

WO2005017174A2 PROCESS FOR THE PURIFICATION OF RECOMBINANT POLYPEPTIDES 审中-公开
标题翻译：重组多肽的纯化方法
公开(公告)号：WO2005017174A2
公开(公告)日：2005-02-24
申请号：PCT/EP2004/009055
申请日：2004-08-12
申请人： SANDOZ AG , STEMPFER, Günter , ALLIGER, Peter , PALMA, Norbert
发明人： STEMPFER, Günter , ALLIGER, Peter , PALMA, Norbert
IPC分类号： C12P21/00
CPC分类号： C07K14/56 , C12P21/02
摘要： This invention relates to a method for purification of a recombinant polypeptide of interest, which polypeptide upon expression has been secreted into the periplasm of a transformed host cell.
摘要翻译：本发明涉及用于纯化感兴趣的重组多肽的方法，所述多肽在表达时已经被分泌到转化的宿主细胞的周质中。

3. 发明申请

WO2005017162A3 EXPRESSION VECTORS, TRANSFORMED HOST CELLS AND FERMENTATION PROCESS FOR THE PRODUCTION OF RECOMBINANT POLYPEPTIDES 审中-公开
公开(公告)号：WO2005017162A3
公开(公告)日：2005-02-24
申请号：PCT/EP2004/009067
申请日：2004-08-12
申请人： SANDOZ AG , WINDISCH, Jörg , SCHOERGENDORFER, Kurt , PALMA, Norbert , KNAUSEDER, Franz , BOEHLING, Hans
发明人： WINDISCH, Jörg , SCHOERGENDORFER, Kurt , PALMA, Norbert , KNAUSEDER, Franz , BOEHLING, Hans
IPC分类号： C12N15/63
摘要： The present invention relates to an expression vector, comprising a polynucleotide encoding a fusion protein comprising the signal sequence of the gac gene of Pseudomonas diminuta and a polypeptide of interest, a prokaryotic host cell transformed with such an expression vector and a process for production of a polypeptide of interest using said host cell and said expression vector.

4. 发明申请

WO2003045996A1 CHROMATOGRAPHIC PURIFICATION OF RECOMBINANT HUMAN ERYTHROPOIETIN 审中-公开
标题翻译：重组人ERYTHROPOIETIN的色谱纯化
公开(公告)号：WO2003045996A1
公开(公告)日：2003-06-05
申请号：PCT/EP2002/013299
申请日：2002-11-26
申请人： BIOCHEMIE GESELLSCHAFT M.B.H. , ALLIGER, Peter , PALMA, Norbert
发明人： ALLIGER, Peter , PALMA, Norbert
IPC分类号： C07K14/505
CPC分类号： C07K14/505
摘要： The invention provides a method for recovering and purifying recombinant human erythropoietin (rhEpo) from a cell culture medium comprising host cells, which method comprises the steps of: (a) removing host cells, cellular constituents and debris from the cell culture medium by centrifugation using a disc stack separator followed by a depeth filtration step to obtain a clarified culture medium supernatant; (b) adjusting the conductivity of the supernatant to 5 mS/cm or less, and a pH of between about 7.0 and 8.0; (c) applying the supernatant from step (b) to a column comprising an anion exchange chromatographic medium, washing the column, eluting the rhEpo from the column, and collecting the peak fraction (s) that contain rhEpo; (d) subjecting the combined peak fractions from step (c) to a reverse phase chromatography step using a polystyrene resin that can be run under medium pressure (
摘要翻译：本发明提供了从包含宿主细胞的细胞培养基中回收和纯化重组人促红细胞生成素（rhEpo）的方法，该方法包括以下步骤：（a）通过离心从细胞培养基中除去宿主细胞，细胞成分和碎片，盘叠分离器，然后进行去过滤步骤以获得澄清的培养基上清液; （b）将上清液的电导率调节至5mS / cm或更低，pH值在约7.0至8.0之间; （c）将来自步骤（b）的上清液施加到包含阴离子交换色谱介质的柱中，洗涤柱，从柱洗脱rhEpo，并收集含有rhEpo的峰分数; （d）使用可在中压（<10巴）下运行并且耐高浓度NaOH（例如Source 30RPC）的聚苯乙烯树脂使步骤（c）的组合峰部分进行反相层析步骤，使用有机溶剂的线性梯度洗脱rhEpo; （e）将含有rhEpo的步骤（d）中洗脱的一种或多种级分施加到包含Q-Seph HP阴离子交换色谱介质的柱上，洗涤柱，并使用线性盐梯度洗脱rhEpo; （f）基于rhEpo的唾液酸化度选择在步骤（e）中洗脱的含有rhEpo的一个或多个级分; 和（g）使用Superdex 75制备级使一种或多种尺寸排阻层析步骤使含有rhEpo的步骤（f）中洗脱的一种或多种级分经脱水以除去潜在的二聚体和较高聚集体; 并收集含有rhEpo的洗脱液。

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