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1. 发明授权

US08183029B2 Replicable expression vector for bacterial expression of a mammalian polypeptide 有权
标题翻译：用于细菌表达哺乳动物多肽的可复制表达载体
公开(公告)号：US08183029B2
公开(公告)日：2012-05-22
申请号：US12727789
申请日：2010-03-19
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12P21/06 , C12N9/00 , C12N9/24 , C12N9/26 , C12N1/20 , C12N15/00 , C07H21/04
CPC分类号： C12N9/2462 , C07K14/52 , C07K14/65 , C12N9/22 , C12N15/70 , C12P21/02
摘要： Refractile particles containing a heterologous polypeptide as an insoluble aggregate are recovered from bacterial periplasm. The process involves culturing bacterial cells so as to express nucleic acid encoding phage lysozyme and nucleic acid encoding the heterologous polypeptide under separate promoters, disrupting the cells mechanically to release the phage lysozyme so as to release refractile particles from the bacterial cellular matrix, and recovering the released refractile particles from the periplasm. Chloroform is not used in any step and the recovery step minimizes co-recovery of cellular debris with the released refractile particles.
摘要翻译：从细菌周质中回收含有异源多肽作为不溶性聚集体的易折射粒子。该方法包括培养细菌细胞，以便在单独的启动子下表达编码噬菌体溶菌酶的核酸和编码异源多肽的核酸，机械破坏细胞以释放噬菌体溶菌酶，从细菌细胞基质中释放可折射颗粒，并回收从周质释放出可折射粒子。氯仿不用于任何步骤，并且回收步骤最小化释放的折射率颗粒的细胞碎片的共同回收。

2. 发明申请

US20110059484A1 Process for producing polypeptides 有权
标题翻译：生产多肽的方法
公开(公告)号：US20110059484A1
公开(公告)日：2011-03-10
申请号：US12586023
申请日：2009-09-16
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12P21/00
CPC分类号： C12N15/67 , C07K16/26 , C07K2317/22 , C12N1/20 , C12N1/32 , C12N1/38 , C12N15/70 , C12P21/00
摘要： A process is described for producing a polypeptide heterologous to E. coli wherein E. coli cells comprising nucleic acid encoding the polypeptide are cultured in a culture medium while feeding to the culture medium a transportable organophosphate, such that the nucleic acid is expressed. The polypeptide is then recovered from the cells.
摘要翻译：描述了用于产生与大肠杆菌异源的多肽的方法，其中包含编码多肽的核酸的大肠杆菌细胞在培养基中培养，同时向培养基中加入可运输的有机磷酸酯，使得表达核酸。然后从细胞中回收多肽。

3. 发明授权

US07294483B2 Process for producing recombinant polypeptides via a glycerol phosphate or sugar phosphate feed 有权
标题翻译：通过甘油磷酸盐或糖磷酸盐饲料生产重组多肽的方法
公开(公告)号：US07294483B2
公开(公告)日：2007-11-13
申请号：US11077272
申请日：2005-03-10
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12N15/09 , C12P21/02
CPC分类号： C12N15/67 , C07K16/26 , C07K2317/22 , C12N1/20 , C12N1/32 , C12N1/38 , C12N15/70 , C12P21/00
摘要： A process is described for producing a polypeptide heterologous to E. coli wherein E. coli cells comprising nucleic acid encoding the polypeptide are cultured in a culture medium while feeding to the culture medium a transportable organophosphate, such that the nucleic acid is expressed. The polypeptide is then recovered from the cells.
摘要翻译：描述了用于产生与大肠杆菌异源的多肽的方法，其中包含编码多肽的核酸的大肠杆菌细胞在培养基中培养，同时向培养基中加入可运输的有机磷酸酯，使得表达核酸。然后从细胞中回收多肽。

4. 发明授权

US06258560B1 Process for bacterial production of polypeptides 有权
标题翻译：细菌生产多肽的方法
公开(公告)号：US06258560B1
公开(公告)日：2001-07-10
申请号：US09607756
申请日：2000-06-29
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12P2100
CPC分类号： C12N9/2462 , C07K14/005 , C07K14/65 , C07K16/2845 , C07K2317/24 , C07K2317/54 , C07K2319/02 , C12N1/06 , C12N9/22 , C12N2795/10222 , C12N2795/10322 , C12P21/02
摘要： Processes are described for recovering heterologous polypeptide from bacterial cells, including the periplasm and cytoplasm. One process involves culturing the bacterial cells, which cells comprise nucleic acid encoding phage lysozyme and nucleic acid encoding a protein that displays DNA-digesting activity, wherein these nucleic acids are linked to a first promoter, and nucleic acid encoding the heterologous polypeptide, which nucleic acid is linked to a second promoter, under certain conditions to produce a broth lysate; and recovering accumulated heterologous polypeptide from the broth lysate. Another process entails culturing bacterial cells that comprise nucleic acid encoding phage lysozyme, gene t, and nucleic acid encoding a protein that displays DNA-digesting activity under the control of a signal sequence for secretion of said DNA-digesting protein, wherein said nucleic acids are linked to one or more promoters, and nucleic acid encoding the heterologous polypeptide and a signal sequence for secretion of the heterologous polypeptide, which nucleic acid encoding the heterologous polypeptide is linked to a another promoter that is inducible, under certain conditions to produce a broth lysate; and recovering accumulated heterologous polypeptide from the broth lysate.
摘要翻译：描述了从细菌细胞（包括周质和细胞质）中回收异源多肽的方法。一种方法包括培养细菌细胞，该细胞包含编码噬菌体溶菌酶的核酸和编码显示DNA消化活性的蛋白质的核酸，其中这些核酸连接到第一启动子，以及编码异源多肽的核酸，该核酸在某些条件下，酸与第二启动子连接以产生肉汤裂解物; 并从肉汤裂解物中回收积累的异源多肽。另一个过程需要培养包含编码噬菌体溶菌酶，基因t的核酸的细菌细胞和编码在用于分泌所述DNA消化蛋白的信号序列控制下显示DNA消化活性的蛋白质的核酸，其中所述核酸是连接到一个或多个启动子，以及编码异源多肽的核酸和用于分泌异源多肽的信号序列，所述异源多肽的编码异源多肽的核酸在某些条件下与可诱导的另一个启动子连接以产生肉汤裂解物 ; 并从肉汤裂解物中回收积累的异源多肽。

5. 发明授权

US07858339B1 Process for bacterial production of polypeptides 有权
标题翻译：细菌生产多肽的方法
公开(公告)号：US07858339B1
公开(公告)日：2010-12-28
申请号：US09422528
申请日：1999-10-21
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12P21/06 , C12N9/00 , C12N9/24 , C12N9/26 , C12N1/20 , C12N15/00 , C07H21/04
CPC分类号： C12N9/2462 , C07K14/52 , C07K14/65 , C12N9/22 , C12N15/70 , C12P21/02
摘要： Refractile particles containing a heterologous polypeptide as an insoluble aggregate are recovered from bacterial periplasm. The process involves culturing bacterial cells so as to express nucleic acid encoding phage lysozyme and nucleic acid encoding the heterologous polypeptide under separate promoters, disrupting the cells mechanically to release the phage lysozyme so as to release refractile particles from the bacterial cellular matrix, and recovering the released refractile particles from the periplasm. Chloroform is not used in any step and the recovery step minimizes co-recovery of cellular debris with the released refractile particles.
摘要翻译：从细菌周质中回收含有异源多肽作为不溶性聚集体的易折射粒子。该方法包括培养细菌细胞，以便在单独的启动子下表达编码噬菌体溶菌酶的核酸和编码异源多肽的核酸，机械破坏细胞以释放噬菌体溶菌酶，从细菌细胞基质中释放可折射颗粒，并回收从周质释放出可折射粒子。氯仿不用于任何步骤，并且回收步骤最小化释放的折射率颗粒的细胞碎片的共同回收。

6. 发明授权

US06180367B2 Process for bacterial production of polypeptides 有权
公开(公告)号：US06180367B2
公开(公告)日：2001-01-30
申请号：US09422712
申请日：1999-10-21
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12P2100
CPC分类号： C12N9/2462 , C07K14/005 , C07K14/65 , C07K16/2845 , C07K2317/24 , C07K2317/54 , C07K2319/02 , C12N1/06 , C12N9/22 , C12N2795/10222 , C12N2795/10322 , C12P21/02
摘要： Processes are described for recovering heterologous polypeptide from bacterial cells, including the periplasm and cytoplasm. One process involves culturing the bacterial cells, which cells comprise nucleic acid encoding phage lysozyme and nucleic acid encoding a protein that displays DNA-digesting activity, wherein these nucleic acids are linked to a first promoter, and nucleic acid encoding the heterologous polypeptide, which nucleic acid is linked to a second promoter, under certain conditions to produce a broth lysate; and recovering accumulated heterologous polypeptide from the broth lysate. Another process entails culturing bacterial cells that comprise nucleic acid encoding phage lysozyme, gene t, and nucleic acid encoding a protein that displays DNA-digesting activity under the control of a signal sequence for secretion of said DNA-digesting protein, wherein said nucleic acids are linked to one or more promoters, and nucleic acid encoding the heterologous polypeptide and a signal sequence for secretion of the heterologous polypeptide, which nucleic acid encoding the heterologous polypeptide is linked to a another promoter that is inducible, under certain conditions to produce a broth lysate; and recovering accumulated heterologous polypeptide from the broth lysate.

7. 发明授权

US08389242B2 Process for producing Apo2L 有权
标题翻译：生产Apo2L的方法
公开(公告)号：US08389242B2
公开(公告)日：2013-03-05
申请号：US12586023
申请日：2009-09-16
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12N15/09 , C12P21/02 , C12P1/04 , C07K14/52
CPC分类号： C12N15/67 , C07K16/26 , C07K2317/22 , C12N1/20 , C12N1/32 , C12N1/38 , C12N15/70 , C12P21/00
摘要： A process is described for producing a polypeptide heterologous to E. coli wherein E. coli cells comprising nucleic acid encoding the polypeptide are cultured in a culture medium while feeding to the culture medium a transportable organophosphate, such that the nucleic acid is expressed. The polypeptide is then recovered from the cells.
摘要翻译：描述了用于产生与大肠杆菌异源的多肽的方法，其中包含编码多肽的核酸的大肠杆菌细胞在培养基中培养，同时向培养基中加入可运输的有机磷酸酯，使得表达核酸。然后从细胞中回收多肽。

8. 发明申请

US20100173389A1 PROCESS FOR BACTERIAL PRODUCTION OF POLYPEPTIDES 有权
标题翻译：多肽生产细菌的方法
公开(公告)号：US20100173389A1
公开(公告)日：2010-07-08
申请号：US12727789
申请日：2010-03-19
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12N1/21 , C12N15/63
CPC分类号： C12N9/2462 , C07K14/52 , C07K14/65 , C12N9/22 , C12N15/70 , C12P21/02
摘要： Refractile particles containing a heterologous polypeptide as an insoluble aggregate are recovered from bacterial periplasm. The process involves culturing bacterial cells so as to express nucleic acid encoding phage lysozyme and nucleic acid encoding the heterologous polypeptide under separate promoters, disrupting the cells mechanically to release the phage lysozyme so as to release refractile particles from the bacterial cellular matrix, and recovering the released refractile particles from the periplasm. Chloroform is not used in any step and the recovery step minimizes co-recovery of cellular debris with the released refractile particles.
摘要翻译：从细菌周质中回收含有异源多肽作为不溶性聚集体的易折射粒子。该方法包括培养细菌细胞，以便在单独的启动子下表达编码噬菌体溶菌酶的核酸和编码异源多肽的核酸，机械破坏细胞以释放噬菌体溶菌酶，从细菌细胞基质中释放可折射颗粒，并回收从周质释放出可折射粒子。氯仿不用于任何步骤，并且回收步骤最小化释放的折射率颗粒的细胞碎片的共同回收。

9. 发明申请

US20120077740A1 Methods for making Apo-2 Ligand using divalent metal ions 审中-公开
标题翻译：使用二价金属离子制备Apo-2配体的方法
公开(公告)号：US20120077740A1
公开(公告)日：2012-03-29
申请号：US12928422
申请日：2010-12-10
申请人： Avi J. Ashkenazi , Sarah Hymowitz , Robert F. Kelley , Iphigenia Koumenis , Woon-Lam Susan Leung , Mark O'Connell , Roger Pai , Zahra Shahrokh , Laura Simmons
发明人： Avi J. Ashkenazi , Sarah Hymowitz , Robert F. Kelley , Iphigenia Koumenis , Woon-Lam Susan Leung , Mark O'Connell , Roger Pai , Zahra Shahrokh , Laura Simmons
IPC分类号： A61K38/19 , C12N15/19 , C12N15/63 , C12N1/21 , B01D15/36 , C07K1/14 , C07K1/18 , C07K1/20 , B01D15/32 , C12P21/02 , C07K14/52
CPC分类号： A61K9/19 , A61K9/0019 , A61K38/00 , A61K47/02 , C07K14/525 , C07K14/70575
摘要： Methods of making Apo-2 ligand and formulations of Apo-2 ligand using divalent metal ions are provided. Such divalent metal ions include zinc and cobalt which improve Apo-2 ligand trimer formation and stability. The crystal structure of Apo-2 ligand is also provided, along with Apo-2 ligand variant polypeptides identified using oligonucleotide-directed mutagenesis.
摘要翻译：提供使用二价金属离子制备Apo-2配体和Apo-2配体制剂的方法。这种二价金属离子包括改善Apo-2配体三聚体形成和稳定性的锌和钴。还提供了Apo-2配体的晶体结构，以及使用寡核苷酸定向突变鉴定的Apo-2配体变体多肽。

10. 发明授权

US07855066B1 Methods for making Apo-2 ligand using divalent metal ions 有权
标题翻译：使用二价金属离子制备Apo-2配体的方法
公开(公告)号：US07855066B1
公开(公告)日：2010-12-21
申请号：US11528948
申请日：2006-09-28
申请人： Avi J. Ashkenazi , Sarah Hymowitz , Robert F. Kelley , Iphigenia Koumenis , Woon-Lam Susan Leung , Mark O'Connell , Roger Pai , Zahra Shahrokh , Laura Simmons
发明人： Avi J. Ashkenazi , Sarah Hymowitz , Robert F. Kelley , Iphigenia Koumenis , Woon-Lam Susan Leung , Mark O'Connell , Roger Pai , Zahra Shahrokh , Laura Simmons
IPC分类号： C12N5/10 , C12N15/12 , C07K14/52
CPC分类号： A61K9/19 , A61K9/0019 , A61K38/00 , A61K47/02 , C07K14/525 , C07K14/70575
摘要： Methods of making Apo-2 ligand and formulations of Apo-2 ligand using divalent metal ions are provided. Such divalent metal ions include zinc and cobalt which improve Apo-2 ligand trimer formation and stability. The crystal structure of Apo-2 ligand is also provided, along with Apo-2 ligand variant polypeptides identified using oligonucleotide-directed mutagenesis.
摘要翻译：提供使用二价金属离子制备Apo-2配体和Apo-2配体制剂的方法。这种二价金属离子包括改善Apo-2配体三聚体形成和稳定性的锌和钴。还提供了Apo-2配体的晶体结构，以及使用寡核苷酸定向突变鉴定的Apo-2配体变体多肽。

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