公开(公告)号：US6051399A

公开(公告)日：2000-04-18

申请号：US934171

申请日：1997-09-19

申请人： Jay S. Stout ,  Bruce E. Partridge ,  Dennis B. Henriksen ,  Barton Holmquist ,  Fred W. Wagner

发明人： Jay S. Stout ,  Bruce E. Partridge ,  Dennis B. Henriksen ,  Barton Holmquist ,  Fred W. Wagner

IPC分类号： C07K1/113 ,  C07K14/60 ,  C07K14/605 ,  C07K14/635 ,  C12N9/88 ,  C12N15/62 ,  C12P21/02 ,  C12P21/06 ,  C12P21/00 ,  C07H21/04 ,  C07K14/46

CPC分类号： C12P21/02 ,  C07K1/113 ,  C07K14/60 ,  C07K14/605 ,  C07K14/635 ,  C12N15/62 ,  C12N9/88 ,  C12P21/06 ,  C07K2319/00 ,  C07K2319/75

摘要： A method for the production of C-terminal amidated recombinant peptides is provided. The method employs a recombinant protein construct having multiple copies of a target peptide linked by intraconnecting peptides. The intraconnecting peptides permit the multicopy construct to be selectively reacted to produce product peptides having a C-terminal .alpha.-carboxamide. A recombinant gene containing a DNA sequence coding for the recombinant protein construct and an expression cassette, an expression vector and a transformed cell including the recombinant gene are also provided.

摘要翻译： 提供了制备C-末端酰胺化重组肽的方法。 该方法采用具有通过连接内肽连接的多肽拷贝的重组蛋白质。 连接肽允许多拷贝构建体选择性地反应以产生具有C-末端α-甲酰胺的产物肽。 还提供了含有编码重组蛋白质构建体的DNA序列和表达盒，表达载体和包含重组基因的转化细胞的重组基因。

公开(公告)号：US06410707B1

公开(公告)日：2002-06-25

申请号：US09750913

申请日：2001-01-02

申请人： Fred W. Wagner ,  Jay S. Stout ,  Dennis B. Henriksen ,  Bruce E. Partridge ,  Bart Holmquist ,  Julie A. Frank

发明人： Fred W. Wagner ,  Jay S. Stout ,  Dennis B. Henriksen ,  Bruce E. Partridge ,  Bart Holmquist ,  Julie A. Frank

IPC分类号： C07H2104

摘要： A process for the recombinant preparation of a calcitonin fragment and the use of the fragment in the preparation of calcitonin and related analogs is provided. The process includes recombinantly forming a fusion protein which includes the calcitonin fragment linked to a carbonic anhydrase. The recombinantly formed fusion protein is subsequently cleaved to produce a polypeptide which includes the calcitonin fragment. A method for producing a calcitonin carba analog which includes condensing a desaminononapeptide with the recombinantly formed calcitonin fragment is also provided.

公开(公告)号：US06251635B1

公开(公告)日：2001-06-26

申请号：US09139819

申请日：1998-08-25

申请人： Fred W. Wagner ,  Jay S. Stout ,  Dennis B. Henriksen ,  Bruce E. Partridge ,  Bart Holmquist ,  Julie A. Frank

发明人： Fred W. Wagner ,  Jay S. Stout ,  Dennis B. Henriksen ,  Bruce E. Partridge ,  Bart Holmquist ,  Julie A. Frank

IPC分类号： C12P2104

CPC分类号： C07K14/585 ,  C07K2319/00 ,  C12N9/88

摘要： A process for the recombinant preparation of a calcitonin fragment and the use of the fragment in the preparation of calcitonin and related analogs is provided. The process includes recombinantly forming a fusion protein which includes the calcitonin fragment linked to a carbonic anhydrase. The recombinantly formed fusion protein is subsequently cleaved to produce a polypeptide which includes the calcitonin fragment. A method for producing a calcitonin carba analog which includes condensing a desaminononapeptide with the recombinantly formed calcitonin fragment is also provided.

摘要翻译： 提供了重组制剂降钙素片段的方法和该片段在制备降钙素和相关类似物中的用途。 该方法包括重组形成融合蛋白，其包括与碳酸酐酶连接的降钙素片段。 随后切割重组形成的融合蛋白以产生包含降钙素片段的多肽。 还提供了一种降钙素carb类似物的制备方法，其包括将脱氨基肽与重组形成的降钙素片段缩合。

公开(公告)号：US5962270A

公开(公告)日：1999-10-05

申请号：US595868

申请日：1996-02-06

申请人： Fred W. Wagner ,  Jay S. Stout ,  Dennis B. Henriksen ,  Bruce E. Partridge ,  Bart Holmquist ,  Julie A. Frank

发明人： Fred W. Wagner ,  Jay S. Stout ,  Dennis B. Henriksen ,  Bruce E. Partridge ,  Bart Holmquist ,  Julie A. Frank

IPC分类号： C07K14/585 ,  C12N9/88 ,  C12N15/16 ,  C12P21/04 ,  C12P21/00 ,  C12P21/06

CPC分类号： C07K14/585 ,  C12N9/88 ,  C07K2319/00

摘要： A process for the recombinant preparation of a calcitonin fragment and the use of the fragment in the preparation of calcitonin and related analogs is provided. The process includes recombinantly forming a fusion protein which includes the calcitonin fragment linked to a carbonic anhydrase. The recombinantly formed fusion protein is subsequently cleaved to produce a polypeptide which includes the calcitonin fragment. A method for producing a calcitonin carba analog which includes condensing a desaminononapeptide with the recombinantly formed calcitonin fragment is also provided.

公开(公告)号：US06461834B1

公开(公告)日：2002-10-08

申请号：US09212663

申请日：1998-12-16

申请人： Dan Dormady ,  Jay S. Stout ,  Daniel J. Strydom ,  Barton Holmquist ,  Fred W. Wagner

发明人： Dan Dormady ,  Jay S. Stout ,  Daniel J. Strydom ,  Barton Holmquist ,  Fred W. Wagner

IPC分类号： C12P2106

CPC分类号： C07K14/605 ,  C12P21/06

摘要： The invention provides a method of producing a polypeptide having a C-terminal &agr;-carboxamide group. It particularly concerns an enzymatic modification of selected substrate polypeptides which result in cleavage of the substrate polypeptide to form a product peptide with a C-terminal arginine residue having an &agr;-carboxamide group (C-terminal “Arg-NH2”). The method includes contacting an aqueous-based solution including (i) ammonia reagent and (ii) the substrate polypeptide with (iii) clostripain.

摘要翻译： 本发明提供了产生具有C-末端α-甲酰胺基团的多肽的方法。 它特别涉及所选择的底物多肽的酶修饰，其导致底物多肽切割以形成具有α-甲酰胺基团（C-末端“Arg-NH 2”）的C-末端精氨酸残基的产物肽。 该方法包括使含有（i）氨试剂和（ii）底物多肽的水性溶液与（iii）clostripain接触。

公开(公告)号：US5532136A

公开(公告)日：1996-07-02

申请号：US96671

申请日：1993-06-22

申请人： Randall R. Carlson ,  Jay S. Stout ,  Dwane E. Wylie ,  Fred W. Wagner ,  Malcolm Riddell

发明人： Randall R. Carlson ,  Jay S. Stout ,  Dwane E. Wylie ,  Fred W. Wagner ,  Malcolm Riddell

IPC分类号： C07K16/00 ,  C07K16/40 ,  C07K16/44 ,  C12Q1/28 ,  G01N33/53 ,  G01N33/543 ,  G01N33/573 ,  G01N33/84

CPC分类号： C07K16/00 ,  C07K16/40 ,  C07K16/44 ,  G01N33/5308 ,  G01N33/573 ,  G01N33/84

摘要： The invention provides method and kits for detecting a metallic cation in a sample of a body fluid. The preferred method and kits include the use of at least two different types of antibodies having different specificities. In the preferred method, the sample of body fluid can be contacted with an effective amount of a capture antibody specific for a naturally occurring polypeptide that can bind the metallic cation to form a first antigen-antibody complex. An effective amount of an antibody specific for an epitope on a metallic cation-naturally occurring polypeptide complex or an antibody specific for a metallic cation is added to the first antigen-antibody complex to form a second antigen-antibody complex. The amount of the metallic cation in the sample of body fluid is determined by detecting the amount of the second antigen-antibody complex.

摘要翻译： 本发明提供了用于检测体液样品中的金属阳离子的方法和试剂盒。 优选的方法和试剂盒包括使用至少两种具有不同特异性的不同类型的抗体。 在优选的方法中，体液样品可以与有效量的天然存在的多肽捕获抗体接触，所述捕获抗体可以结合金属阳离子以形成第一抗原 - 抗体复合物。 将金属阳离子天然存在的多肽复合物或金属阳离子特异性抗体的表位的特异性抗体的有效量加入到第一抗原 - 抗体复合物中以形成第二抗原 - 抗体复合物。 通过检测第二抗原 - 抗体复合物的量来确定体液样品中金属阳离子的量。

公开(公告)号：US5620856A

公开(公告)日：1997-04-15

申请号：US469489

申请日：1995-06-06

申请人： Randall R. Carlson ,  Jay S. Stout ,  Dwane E. Wylie ,  Fred W. Wagner ,  Malcolm Riddell

发明人： Randall R. Carlson ,  Jay S. Stout ,  Dwane E. Wylie ,  Fred W. Wagner ,  Malcolm Riddell

IPC分类号： C07K16/00 ,  C07K16/40 ,  C07K16/44 ,  C12Q1/28 ,  G01N33/53 ,  G01N33/543 ,  G01N33/573 ,  G01N33/84

CPC分类号： C07K16/00 ,  C07K16/40 ,  C07K16/44 ,  G01N33/5308 ,  G01N33/573 ,  G01N33/84

摘要： The invention provides method and kits for detecting a metallic cation in a sample of a body fluid. The preferred method and kits include the use of at least two different types of antibodies having different specificities. In the preferred method, the sample of body fluid can be contacted with an effective amount of a capture antibody specific for a naturally occurring polypeptide that can bind the metallic cation to form a first antigen-antibody complex. An effective amount of an antibody specific for an epitope on a metallic cation-naturally occurring polypeptide complex or an antibody specific for a metallic cation is added to the first antigen-antibody complex to form a second antigen-antibody complex. The amount of the metallic cation in the sample of body fluid is determined by detecting the amount of the second antigen-antibody complex.

摘要翻译： 本发明提供了用于检测体液样品中的金属阳离子的方法和试剂盒。 优选的方法和试剂盒包括使用至少两种具有不同特异性的不同类型的抗体。 在优选的方法中，体液样品可以与有效量的天然存在的多肽捕获抗体接触，所述捕获抗体可以结合金属阳离子以形成第一抗原 - 抗体复合物。 将金属阳离子天然存在的多肽复合物或金属阳离子特异性抗体的表位的特异性抗体的有效量加入到第一抗原 - 抗体复合物中以形成第二抗原 - 抗体复合物。 通过检测第二抗原 - 抗体复合物的量来确定体液样品中金属阳离子的量。

公开(公告)号：US08003362B2

公开(公告)日：2011-08-23

申请号：US10997697

申请日：2004-11-24

申请人： Fred W. Wagner ,  Peng Luan ,  Yuannan Xia ,  Barton Holmquist

发明人： Fred W. Wagner ,  Peng Luan ,  Yuannan Xia ,  Barton Holmquist

IPC分类号： C12N9/48 ,  C12N1/20 ,  C12N15/74 ,  C07K1/00

CPC分类号： C12N9/50 ,  C12N15/67

摘要： The invention provides methods and nucleic acid constructs to express clostripain. The source of the coding region for recombinantly expressed clostripain is Clostridium histolyticum.

摘要翻译： 本发明提供了表达clostripain的方法和核酸构建体。 用于重组表达的clostripain的编码区的来源是溶组织梭菌（Clostridium histolyticum）。

公开(公告)号：US20080153129A1

公开(公告)日：2008-06-26

申请号：US11944165

申请日：2007-11-21

申请人： Fred W. Wagner ,  Peng Luan ,  Yuannan Xia ,  Daniel Strydom ,  Edwin H. Merrifield ,  Mary J. Bossard ,  Barton Holmquist ,  Jin Seog Seo

发明人： Fred W. Wagner ,  Peng Luan ,  Yuannan Xia ,  Daniel Strydom ,  Edwin H. Merrifield ,  Mary J. Bossard ,  Barton Holmquist ,  Jin Seog Seo

IPC分类号： C12P21/02

CPC分类号： C07K14/605 ,  C12N9/52 ,  C12P21/06

摘要： The invention provides methods for making peptides from a polypeptide containing at least one copy of the peptide using clostripain to excise the peptide from the polypeptide. The methods enable the use of a single, highly efficient enzymatic cleavage to produce any desired peptide sequence.

摘要翻译： 本发明提供了使用clostripain从含有至少一个拷贝的肽的多肽制备肽从多肽中切下肽的方法。 这些方法使得能够使用单一的高效酶裂解产生任何所需的肽序列。

公开(公告)号：US20120156719A1

公开(公告)日：2012-06-21

申请号：US12354053

申请日：2009-01-15

申请人： Fred W. Wagner ,  Peng Luan ,  Yuannan Xia ,  Daniel Strydom ,  Edwin H. Merrifield ,  Mary J. Bossard ,  Barton Holmquist ,  Jin Seog Seo

发明人： Fred W. Wagner ,  Peng Luan ,  Yuannan Xia ,  Daniel Strydom ,  Edwin H. Merrifield ,  Mary J. Bossard ,  Barton Holmquist ,  Jin Seog Seo

IPC分类号： C12P21/06

CPC分类号： C07K14/605 ,  C12N9/52 ,  C12P21/06

摘要： The invention provides methods for making peptides from a polypeptide containing at least one copy of the peptide using clostripain to excise the peptide from the polypeptide. The methods enable the use of a single, highly efficient enzymatic cleavage to produce any desired peptide sequence.

摘要翻译： 本发明提供了使用clostripain从含有至少一个拷贝的肽的多肽制备肽从多肽中切下肽的方法。 这些方法使得能够使用单一的高效酶裂解产生任何所需的肽序列。