公开(公告)号：US5656456A

公开(公告)日：1997-08-12

申请号：US457166

申请日：1995-06-01

申请人： Jay Stout ,  Fred W. Wagner ,  Thomas R. Coolidge ,  Bart Holmquist

发明人： Jay Stout ,  Fred W. Wagner ,  Thomas R. Coolidge ,  Bart Holmquist

IPC分类号： C07K1/00 ,  C07K1/107 ,  C07K1/12 ,  C07K5/11 ,  C07K7/18 ,  C07K14/47 ,  C07K14/475 ,  C12N15/62 ,  C12P21/00

CPC分类号： C12N15/62 ,  C07K1/003 ,  C07K1/006 ,  C07K1/1072 ,  C07K1/1075 ,  C07K1/12 ,  C07K14/4716 ,  C07K14/475 ,  C07K5/1019 ,  C07K7/18 ,  C07K2319/00 ,  C07K2319/50 ,  C07K2319/75 ,  Y02P20/55

摘要： The invention provides for a chemical method for preparing a recombinant single copy polypeptide or a portion thereof with a modified terminal amino acid .alpha.-carbon reactive group selected from the group consisting of N-terminal .alpha.-amine, C-terminal .alpha.-carboxyl, and a combination thereof. The steps of the method involve forming the recombinant single copy polypeptide or a portion thereof so that the single copy polypeptide is protected with one or more biologically added protecting groups at the N-terminal .alpha.-amine, C-terminal .alpha.-carboxyl. The recombinant single copy polypeptide can then be reacted with up to three chemical protecting agents to selectively protect reactive side chain groups and thereby prevent side chain groups from being modified. The recombinant single copy polypeptide can be cleaved with at least one cleavage reagent specific for the biological protecting group to form an unprotected terminal amino acid .alpha.-carbon reactive group. The unprotected terminal amino acid .alpha.-carbon reactive group is modified with at least one chemical modifying agent. The side chain protected terminally modified single copy polypeptide is then deprotected at the side chain groups to form a terminally modified recombinant single copy polypeptide. The number and sequence of steps in the method can be varied to achieve selective modification at the N- and/or C-terminal amino acid of a recombinantly produced polypeptide.

摘要翻译： 本发明提供了一种用于制备重组单拷贝多肽或其部分的化学方法，其具有修饰的末端氨基酸α-碳反应性基团，其选自N-末端α-胺，C-末端α-羧基和 其组合。 该方法的步骤包括形成重组单拷贝多肽或其部分，使得单拷贝多肽在N-末端α-胺C-末端α-羧基上用一个或多个生物学添加的保护基进行保护。 然后可将重组单拷贝多肽与多达三种化学保护剂反应，以选择性保护反应性侧链基团，从而防止侧链基团被修饰。 重组单拷贝多肽可以用至少一种对生物保护基特异性的切割试剂进行切割以形成未保护的末端氨基酸α-碳反应性基团。 未保护的末端氨基酸α-碳反应性基团用至少一种化学改性剂进行改性。 然后将侧链保护的末端修饰的单拷贝多肽在侧链基团脱保护以形成末端修饰的重组单拷贝多肽。 可以改变方法中的步骤数目和顺序，以实现重组产生的多肽的N-和/或C-末端氨基酸的选择性修饰。

公开(公告)号：US5635371A

公开(公告)日：1997-06-03

申请号：US294434

申请日：1994-08-23

申请人： Jay Stout ,  Fred W. Wagner ,  Thomas R. Coolidge ,  Bart Holmquist

发明人： Jay Stout ,  Fred W. Wagner ,  Thomas R. Coolidge ,  Bart Holmquist

IPC分类号： C12N15/09 ,  C07K1/00 ,  C07K1/06 ,  C07K1/107 ,  C07K1/113 ,  C07K1/12 ,  C07K5/11 ,  C07K7/18 ,  C07K14/47 ,  C07K14/475 ,  C07K14/60 ,  C07K19/00 ,  C12N15/62 ,  C12P21/02 ,  C12P21/00

CPC分类号： C07K14/60 ,  C07K1/003 ,  C07K1/006 ,  C07K1/1072 ,  C07K1/1075 ,  C07K1/12 ,  C07K14/4716 ,  C07K14/475 ,  C07K5/1019 ,  C07K7/18 ,  C12N15/62 ,  C07K2319/00 ,  C07K2319/50 ,  C07K2319/75 ,  Y02P20/55

摘要： The invention provides for a chemical method for preparing a recombinant single copy polypeptide or a portion thereof with a modified terminal amino acid .alpha.-carbon reactive group selected from the group consisting of N-terminal .alpha.-amine, C-terminal .alpha.-carboxyl, and a combination thereof. The steps of the method involve forming the recombinant single copy polypeptide or a portion thereof so that the single copy polypeptide is protected with one or more biologically added protecting groups at the N-terminal .alpha.-amine, C-terminal .alpha.-carboxyl. The recombinant single copy polypeptide can then be reacted with up to three chemical protecting agents to selectively protect reactive side chain groups and thereby prevent side chain groups from being modified. The recombinant single copy polypeptide can be cleaved with at least one cleavage reagent specific for the biological protecting group to form an unprotected terminal amino acid .alpha.-carbon reactive group. The unprotected terminal amino acid .alpha.-carbon reactive group is modified with at least one chemical modifying agent. The side chain protected terminally modified single copy polypeptide is then deprotected at the side chain groups to form a terminally modified recombinant single copy polypeptide. The number and sequence of steps in the method can be varied to achieve selective modification at the N- and/or C-terminal amino acid of a recombinantly produced polypeptide.

摘要翻译： 本发明提供了一种用于制备重组单拷贝多肽或其部分的化学方法，其具有修饰的末端氨基酸α-碳反应性基团，其选自N-末端α-胺，C-末端α-羧基和 其组合。 该方法的步骤包括形成重组单拷贝多肽或其部分，使得单拷贝多肽在N-末端α-胺C-末端α-羧基上用一个或多个生物学添加的保护基进行保护。 然后可将重组单拷贝多肽与多达三种化学保护剂反应，以选择性保护反应性侧链基团，从而防止侧链基团被修饰。 重组单拷贝多肽可以用至少一种对生物保护基特异性的切割试剂进行切割以形成未保护的末端氨基酸α-碳反应性基团。 未保护的末端氨基酸α-碳反应性基团用至少一种化学改性剂进行改性。 然后将侧链保护的末端修饰的单拷贝多肽在侧链基团脱保护以形成末端修饰的重组单拷贝多肽。 可以改变方法中的步骤数目和顺序，以实现重组产生的多肽的N-和/或C-末端氨基酸的选择性修饰。

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.