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1. 发明授权

US06756484B1 IGF-I purification process 有权
标题翻译： IGF-I纯化方法
公开(公告)号：US06756484B1
公开(公告)日：2004-06-29
申请号：US09614947
申请日：2000-07-12
申请人： Russell A. Brierley , Joan N. Abrams , John M. Hanson , Francis C. Maslanka , Cynthia Cowgill , Luis G. Juarbe-Osorio , Patricio Riquelme , Glenn Dorin
发明人： Russell A. Brierley , Joan N. Abrams , John M. Hanson , Francis C. Maslanka , Cynthia Cowgill , Luis G. Juarbe-Osorio , Patricio Riquelme , Glenn Dorin
IPC分类号： C07K14475
CPC分类号： C07K14/65 , G01N30/02 , B01D15/422
摘要： Disclosed is an improved process for obtaining purified, monomeric, intact, correctly-folded insulin-like growth factor-I (also known as somatomedin-C). The improvements, consisting primarily of the addition of an IGF-I unfolding/refolding step and the substitution of a reverse phase chromatography step for a gel filtration chromatography step result in a three-fold increase in final yield. The process includes the following steps, in order: first cation exchange, unfolding/refolding, hydrophobic interaction chromatography, second cation exchange, and reverse phase chromatography.
摘要翻译：公开了用于获得纯化的，单体的，完整的，正确折叠的胰岛素样生长因子-I（也称为生长因子C）的改进方法。主要由IGF-I展开/重折叠步骤的添加和用于凝胶过滤层析步骤的反相色谱步骤的替代的改进导致最终产率增加三倍。该方法包括以下步骤：首先进行阳离子交换，展开/重折叠，疏水相互作用层析，第二次阳离子交换和反相色谱法。

2. 发明授权

US07193042B1 Methods for purifying authentic IGF from yeast hosts 失效
标题翻译：从酵母宿主纯化正确的IGF的方法
公开(公告)号：US07193042B1
公开(公告)日：2007-03-20
申请号：US08477984
申请日：1995-06-07
申请人： Cynthia Cowgill , Luis Juarbe-Osorio , Patricio Riquelme , Glenn Dorin , Christopher M. Bussineau , Robert D. Kudrna , Carl Scandella , Russell A. Brierley , Joan N. Abrams , John M. Hanson , Francis C. Maslanka
发明人： Cynthia Cowgill , Luis Juarbe-Osorio , Patricio Riquelme , Glenn Dorin , Christopher M. Bussineau , Robert D. Kudrna , Carl Scandella , Russell A. Brierley , Joan N. Abrams , John M. Hanson , Francis C. Maslanka
IPC分类号： C07K1/18 , C07K1/20 , C07K1/22
CPC分类号： C07K14/65
摘要： Methods for purifying authentic, properly folded IGF polypeptides from yeast transformants are disclosed. The methods include a refolding step and provide for high yields of authentic IGF from a variety of yeast strains.
摘要翻译：公开了从酵母转化体纯化正确的，适当折叠的IGF多肽的方法。所述方法包括重折叠步骤，并提供来自各种酵母菌株的高产量的正确的IGF。

3. 发明授权

US07071313B1 Methods of purifying authentic IGF from yeast hosts 失效
标题翻译：从酵母宿主纯化正常的IGF的方法
公开(公告)号：US07071313B1
公开(公告)日：2006-07-04
申请号：US09564414
申请日：2000-05-02
申请人： Cynthia Cowgill , Luis Juarbe-Osorio , Patricio Riquelme , Glenn Dorin , Christopher M. Bussineau , Robert D Kudrna , Asuman G. Ozturk , Carl Scandella , Russell A. Brierley , Joan N. Abrams , John M. Hanson , Francis C. Maslanka
发明人： Cynthia Cowgill , Luis Juarbe-Osorio , Patricio Riquelme , Glenn Dorin , Christopher M. Bussineau , Robert D Kudrna , Asuman G. Ozturk , Carl Scandella , Russell A. Brierley , Joan N. Abrams , John M. Hanson , Francis C. Maslanka
IPC分类号： C07K1/18
CPC分类号： C07K14/65
摘要： Methods for purifying authentic, properly folded IGF polypeptides from yeast transformants are disclosed. The methods include a refolding step and provide for high yields of authentic IGF from a variety of yeast strains.
摘要翻译：公开了从酵母转化体纯化正确的，适当折叠的IGF多肽的方法。所述方法包括重折叠步骤，并提供来自各种酵母菌株的高产量的正确的IGF。

4. 发明授权

US5650496A IGF-I purification process 失效
标题翻译： IGF-I纯化方法
公开(公告)号：US5650496A
公开(公告)日：1997-07-22
申请号：US422436
申请日：1995-04-14
申请人： Russell A. Brierley , Joan N. Abrams , John M. Hanson , Francis C. Maslanka
发明人： Russell A. Brierley , Joan N. Abrams , John M. Hanson , Francis C. Maslanka
IPC分类号： C12N15/09 , C07K1/18 , C07K1/20 , C07K1/36 , C07K14/65 , C12N15/18 , C12P21/00 , C12R1/84 , G01N30/02 , C07K14/475
CPC分类号： C07K14/65 , G01N30/02
摘要： Disclosed is an improved process for obtaining purified, monomeric, intact, correctly-folded insulin-like growth factor-I (also known as somatomedin-C). The improvements, consisting primarily of the addition of an IGF-I unfolding/refolding step and the substitution of a reverse phase chromatography step for a gel filtration chromatography step result in a three-fold increase in final yield. The process includes the following steps, in order: first cation exchange, unfolding/refolding, hydrophobic interaction chromatography, second cation exchange, and reverse phase chromatography.
摘要翻译：公开了用于获得纯化的，单体的，完整的，正确折叠的胰岛素样生长因子-I（也称为生长因子C）的改进方法。主要由IGF-I展开/重折叠步骤的添加和用于凝胶过滤层析步骤的反相色谱步骤的替代的改进导致最终产率增加三倍。该方法包括以下步骤：首先进行阳离子交换，展开/重折叠，疏水相互作用层析，第二次阳离子交换和反相色谱法。

5. 发明授权

US06207806B1 IGF-I purification process 失效
标题翻译： IGF-I纯化方法
公开(公告)号：US06207806B1
公开(公告)日：2001-03-27
申请号：US08851162
申请日：1997-05-05
申请人： Russell A. Brierley , Joan N. Abrams , John M. Hanson , Francis C. Maslanka
发明人： Russell A. Brierley , Joan N. Abrams , John M. Hanson , Francis C. Maslanka
IPC分类号： C07K14475
CPC分类号： C07K14/65 , G01N30/02 , B01D15/422
摘要： Disclosed is an improved process for obtaining purified, monomeric, intact, correctly-folded insulin-like growth factor-I (also known as somatomedin-C). The improvements, consisting primarily of the addition of an IGF-I unfolding/refolding step and the substitution of a reverse phase chromatography step for a gel filtration chromatography step result in a three-fold increase in final yield. The process includes the following steps, in order: first cation exchange, unfolding/refolding, hydrophobic interaction chromatography, second cation exchange, and reverse phase chromatography.
摘要翻译：公开了用于获得纯化的，单体的，完整的，正确折叠的胰岛素样生长因子-I（也称为生长因子C）的改进方法。主要由IGF-I展开/重折叠步骤的添加和用于凝胶过滤层析步骤的反相色谱步骤的替代的改进导致最终产率增加三倍。该方法包括以下步骤：首先进行阳离子交换，展开/重折叠，疏水相互作用层析，第二次阳离子交换和反相色谱法。

6. 发明授权

US5707828A Expression of human serum albumin in Pichia pastoris 失效
标题翻译：人血清白蛋白在毕赤酵母中的表达
公开(公告)号：US5707828A
公开(公告)日：1998-01-13
申请号：US433037
申请日：1995-05-03
申请人： Kotikanyadan Sreekrishna , Juerg F. Tschopp , Gregory P. Thill , Russell A. Brierley , Kathryn A. Barr
发明人： Kotikanyadan Sreekrishna , Juerg F. Tschopp , Gregory P. Thill , Russell A. Brierley , Kathryn A. Barr
IPC分类号： C12N15/09 , C07K14/765 , C12N1/15 , C12N1/19 , C12N15/14 , C12N15/81 , C12P21/02 , C12R1/84 , C12P21/06 , C12N15/00 , C12N15/63
CPC分类号： C12N15/815 , C07K14/765
摘要： The present invention is directed to expression cassettes comprising a 5' regulatory region from at least one of the Pichia pastoris AOXI gene, p40 gene, DASI gene or HIS4 gene, operably linked to an HSA structural gene including the HSA signal sequence. The HSA structural gene has a translational start codon within 0 to 11 deoxyribonucleotides from the 5' end of the HSA structural gene and is operably linked to a 3' termination sequence. Further, the adenine and thymine content of the intervening deoxyribonucleotides is in the range from about 55 to about 64%. The expression cassette may be an autonomously replicating vector or an integrative vector. Other embodiments of the present invention include Pichia pastoris strains transformed with the HSA expression cassettes and processes for secretion of HSA using the expression cassettes.
摘要翻译：本发明涉及包含与包含HSA信号序列的HSA结构基因可操作地连接的至少一种巴斯德毕赤酵母AOXI基因，p40基因，DASI基因或HIS4基因的5'调节区的表达盒。 HSA结构基因具有来自HSA结构基因5'末端的0至11个脱氧核糖核苷酸之间的翻译起始密码子，并且可操作地连接到3'终止序列。此外，介入的脱氧核糖核苷酸的腺嘌呤和胸腺嘧啶含量在约55至约64％的范围内。表达盒可以是自主复制载体或整合载体。本发明的其他实施方案包括用HSA表达盒转化的巴斯德毕赤酵母菌株和使用表达盒分泌HSA的方法。

7. 发明授权

US5612198A Production of insulin-like growth factor-1 in methylotrophic yeast cells 失效
标题翻译：在甲基营养酵母细胞中生产胰岛素样生长因子-1
公开(公告)号：US5612198A
公开(公告)日：1997-03-18
申请号：US308196
申请日：1994-09-19
申请人： Russell A. Brierley , Geneva R. Davis , Gregory C. Holtz , Martin A. Gleeson , Bradley D. Howard
发明人： Russell A. Brierley , Geneva R. Davis , Gregory C. Holtz , Martin A. Gleeson , Bradley D. Howard
IPC分类号： C07K14/65 , C12N15/81 , C12N15/18 , C12N1/19
CPC分类号： C12N15/815 , C07K14/65 , C07K2319/02
摘要： Insulin-like growth factor-1 (IGF-1), a naturally occurring, relatively short, single chain polypeptide, is prepared by growing methylotrophic yeast transformants containing in their genome at least one copy of DNA encoding IGF-1, in operational linkage with DNA encoding a signal sequence, which is effective for directing secretion of proteins from the host cells and which also includes the proteolytic processing site lys-arg and may include one or more glu-ala sequences. In preferred embodiments the signal sequence is the S. cerevisiae alpha mating factor pre-pro sequence. Expression of both the DNA encoding IGF-1 and the pre-pro signal sequence is regulated by a promoter region derived from a methanol responsive gene of a methylotrophic yeast. DNA constructs and recombinant methylotrophic yeast strains used for the expression and secretion of IGF-1 are also provided. For preferred embodiments, protease deficient Pichia pastoris strains are provided.
摘要翻译：胰岛素样生长因子-1（IGF-1）是一种天然存在的相对较短的单链多肽，通过生长含有其基因组的含有编码IGF-1的DNA的至少一个拷贝的甲基营养酵母转化体，与编码信号序列的DNA，其对于引导来自宿主细胞的蛋白质的分泌是有效的，并且还包括蛋白水解加工位点lys-arg，并且可以包括一个或多个葡糖醛酸序列。在优选实施方案中，信号序列是酿酒酵母α交配因子前序序列。编码IGF-1和前一信号序列的DNA的表达由来自甲基营养酵母的甲醇应答基因的启动子区域调节。还提供了用于IGF-1的表达和分泌的DNA构建体和重组甲基营养酵母菌株。对于优选的实施方案，提供了蛋白酶缺乏的巴斯德毕赤酵母菌株。

8. 发明授权

US5231178A Method for the purification of intact, correctly-folded insulin-like growth factor-1 失效
标题翻译：纯化，正确折叠的胰岛素样生长因子-1的方法
公开(公告)号：US5231178A
公开(公告)日：1993-07-27
申请号：US785171
申请日：1991-10-28
申请人： Gregory C. Holtz , Russell A. Brierley
发明人： Gregory C. Holtz , Russell A. Brierley
IPC分类号： C07K1/14 , C07K1/20 , C07K14/00 , C07K14/575 , C07K14/65 , C12N1/19 , C12N15/18 , C12P21/00 , C12R1/84
CPC分类号： C07K14/65 , C07K1/20
摘要： Method for the recovery and purification of intact, correctly-folded, monomeric insulin-like growth factor-1 peptide from large volumes of IGF-1-containing medium are described, comprising a series of adsorption-desorption steps employing a combination of cation exchange and hydrophobic interaction adsorbents. Product IGF-1 peptide is highly purified and suitable for use in a variety of clinical applications.
摘要翻译：描述了从大量含IGF-1的培养基中回收和纯化完整的，正确折叠的单体胰岛素样生长因子-1肽的方法，其包括一系列吸附 - 解吸步骤，其采用阳离子交换和疏水相互作用吸附剂。产品IGF-1肽是高度纯化的，适用于各种临床应用。

9. 发明授权

US06723844B1 Preparation of K-252a 有权
标题翻译： K-252a的制备
公开(公告)号：US06723844B1
公开(公告)日：2004-04-20
申请号：US10375413
申请日：2003-02-27
申请人： Navayath Shobana , John C. Strong , Mark W. Tubergen , Russell A. Brierley
发明人： Navayath Shobana , John C. Strong , Mark W. Tubergen , Russell A. Brierley
IPC分类号： C07D49818
CPC分类号： C07D498/22
摘要： The present invention provides a process for the synthesis of K-252a and intermediates useful in the process.
摘要翻译：本发明提供了合成K-252a的方法和在该方法中有用的中间体。

10. 发明授权

US5521086A Secretion sequence for the production of a heterologous protein in yeast 失效
标题翻译：在酵母中产生异源蛋白质的分泌序列
公开(公告)号：US5521086A
公开(公告)日：1996-05-28
申请号：US122889
申请日：1993-09-16
申请人： Richard W. Scott , Russell A. Brierley , David S. Howland
发明人： Richard W. Scott , Russell A. Brierley , David S. Howland
IPC分类号： C12N15/09 , C07K14/39 , C07K14/65 , C07K19/00 , C12N1/11 , C12N1/19 , C12N9/16 , C12N15/81 , C12P21/02 , C12R1/645 , C12N1/21 , C12N15/62
CPC分类号： C12N15/81 , C07K14/65 , C07K2319/00 , C07K2319/02
摘要： The invention generally features a novel recombinant DNA molecule useful for producing a heterologous protein, e.g., insulin-like growth factor-I, in yeast. The DNA molecule includes the following sequences joined in frame and in the following 5' to 3' progression: a sequence encoding a signal peptide of a yeast secreted protein; a sequence encoding at least 5 amino acids of the N-terminal region of a mature acid phosphatase protein, a sequence encoding a heterologous protein. Preferably the DNA molecule also encodes a spacer element that includes a proteolytic cleavage site.
摘要翻译：本发明通常具有用于在酵母中产生异源蛋白质（例如胰岛素样生长因子-I）的新型重组DNA分子。 DNA分子包括以下框架连接的以下5'至3'进程的序列：编码酵母分泌蛋白质的信号肽的序列; 编码成熟酸性磷酸酶蛋白质N末端区域至少5个氨基酸的序列，编码异源蛋白质的序列。优选地，DNA分子还编码包含蛋白水解切割位点的间隔元件。

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