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1. 发明授权

US5741686A Exopeptidase catalyzed site-specific bonding of supports, labels and bioactive agents to proteins 失效
标题翻译：外肽酶催化载体，标记和生物活性剂对蛋白质的位点特异性结合
公开(公告)号：US5741686A
公开(公告)日：1998-04-21
申请号：US316810
申请日：1994-10-03
申请人： Fred W. Wagner , Thomas R. Coolidge , Dwane E. Wylie , Sheldon M. Schuster , William Lewis , Jay Stout
发明人： Fred W. Wagner , Thomas R. Coolidge , Dwane E. Wylie , Sheldon M. Schuster , William Lewis , Jay Stout
IPC分类号： C07K17/06 , C07K1/107 , C07K14/575 , C07K16/00 , C07K16/40 , C07K17/00 , C07K19/00 , C12N11/00 , C12N11/02 , C12N11/06 , G01N33/532 , G01N33/543 , G01N33/58 , C12N9/96 , C07K1/13 , C08H1/00
CPC分类号： C07K1/1077 , C07K16/40 , C07K17/00 , C12N11/00 , G01N33/532 , G01N33/543 , G01N33/58 , Y10S530/811 , Y10S530/812 , Y10S530/815 , Y10S530/816
摘要： A method is provided for preparing a labeled protein, immobilized protein or protein-bioactive agent composition by attaching a label, support or bioactive agent to a protein by exopeptidase catalysis at a site that is remote from the active site of the protein. More specifically, an amine or alcohol group of an amino acid, amine or alcohol nucleophile is reacted by exopeptidase catalysis with a C-terminus carboxylic acid group of a protein such as an antibody, enzyme or hormone to couple the nucleophile to the protein to form an adduct, and the adduct is bound to an auxiliary substance such as a support, label or bioactive agent or its combination with a linker arm by reacting a reactive substituent of the nucleophile with a reactive group of the auxiliary substance. Alternatively, the nucleophile is bound to the auxiliary substance or its combination with a linker arm to form an intermediate, and the intermediate is coupled by exopeptidase catalysis to the protein. The exopeptidase may be a serine or cysteine exocarboxypeptidase.
摘要翻译：提供了一种用于通过在远离蛋白质活性位点的位点处通过外肽酶催化将标记物，载体或生物活性剂附着到蛋白质来制备标记的蛋白质，固定的蛋白质或蛋白质 - 生物活性剂组合物的方法。更具体地，氨基酸，胺或醇亲核试剂的胺或醇基团通过外肽酶催化与诸如抗体，酶或激素的蛋白质的C末端羧酸基团反应，以将亲核体偶联到蛋白质上形成加合物，并且加合物通过将亲核试剂的反应性取代基与辅助物质的反应性基团反应而与辅助物质例如载体，标记或生物活性剂或其与连接臂的组合结合。或者，亲核试剂与辅助物质或其与连接臂的组合结合以形成中间体，并且中间体通过外肽酶催化与蛋白质偶联。外肽酶可以是丝氨酸或半胱氨酸外切羧肽酶。

2. 发明授权

US5234820A Exopeptidase catalyzed site-specific bonding of supports, labels and bioactive agents to proteins 失效
标题翻译：外肽酶催化载体，标记和生物活性剂对蛋白质的位点特异性结合
公开(公告)号：US5234820A
公开(公告)日：1993-08-10
申请号：US545915
申请日：1990-06-28
申请人： Fred W. Wagner , Thomas R. Coolidge , Sheldon M. Schuster , Jay Stout , Dwane E. Wylie , Klaus Breddam , William Lewis
发明人： Fred W. Wagner , Thomas R. Coolidge , Sheldon M. Schuster , Jay Stout , Dwane E. Wylie , Klaus Breddam , William Lewis
IPC分类号： C07K1/107 , C07K16/40 , C07K17/00 , C12N11/00 , G01N33/532 , G01N33/543 , G01N33/58
CPC分类号： C12N11/00 , C07K1/1077 , C07K16/40 , C07K17/00 , G01N33/532 , G01N33/543 , G01N33/58 , Y10S530/816
摘要： An auxiliary substance such as a label, support, or bioactive agent is attached to a protein at a site that is remote from the active site of the protein by the use of exopeptidase and a nucleophile which is an amino acid, amino acid derivative, amine or alcohol. In one embodiment, the nucleophile is attached to the carboxy terminus of a protein by catalysis with exopeptidase to form an adduct and then the adduct or its combination with a linker arm is bound to the auxiliary substance. In another embodiment, the auxiliary substance or its combination with a linker arm is bound to the nucleophile to form an intermediate substance which is then coupled by catalysis with exopeptidase to the carboxy terminus of a protein.
摘要翻译：辅助物质如标记物，载体或生物活性剂通过使用外肽酶和亲核试剂（氨基酸，氨基酸衍生物，胺）与远离蛋白质活性位点的位点连接到蛋白质上或酒精。在一个实施方案中，亲核试剂通过使用外肽酶的催化与蛋白质的羧基末端连接以形成加合物，然后加合物或其与连接臂的组合与辅助物质结合。在另一个实施方案中，辅助物质或其与连接臂的组合与亲核试剂结合以形成中间体物质，然后通过与外肽酶的催化偶联至蛋白质的羧基末端。

3. 发明授权

US5279954A Exopeptidase catalyzed site-specific bonding of supports, labels and bioactive agents to proteins 失效
标题翻译：外肽酶催化载体，标记和生物活性剂对蛋白质的位点特异性结合
公开(公告)号：US5279954A
公开(公告)日：1994-01-18
申请号：US375138
申请日：1989-06-30
申请人： Fred W. Wagner , Thomas R. Coolidge , Dwane E. Wylie , Sheldon M. Schuster , William Lewis , Jay Stout
发明人： Fred W. Wagner , Thomas R. Coolidge , Dwane E. Wylie , Sheldon M. Schuster , William Lewis , Jay Stout
IPC分类号： C07K17/06 , C07K1/107 , C07K14/575 , C07K16/00 , C07K16/40 , C07K17/00 , C07K19/00 , C12N11/00 , C12N11/02 , C12N11/06 , G01N33/532 , G01N33/543 , G01N33/58 , C12N11/14 , G01N33/544 , G01N33/551
CPC分类号： C07K1/1077 , C07K16/40 , C07K17/00 , C12N11/00 , G01N33/532 , G01N33/543 , G01N33/58 , Y10S530/811 , Y10S530/812 , Y10S530/815 , Y10S530/816
摘要： The invention provides a means for attaching a label, support or bioactive agent to a protein with an exopeptidase at a site that is remote from the active site of the protein. More specifically the invention is directed to a method for the attachment of an amino acid, amine and alcohol nucleophile to the carboxyl terminus of a protein. In one embodiment, a labeled nucleophile is attached to a protein such as an antibody. In other embodiments, the invention is directed to a method for the attachment of a protein to an immobilization support and to a method for the attachment of a bioactive agent to a protein.
摘要翻译：本发明提供了一种在远离蛋白质活性位点的位点附着标记物，载体或生物活性剂到蛋白质的方法。更具体地，本发明涉及将氨基酸，氨基酸衍生物，胺和醇亲核试剂连接到蛋白质的羧基末端的方法。在一个实施方案中，使用外肽酶将标记的亲核试剂连接到蛋白质例如抗体。在其它实施方案中，本发明涉及将蛋白质与固定化载体连接的方法以及将生物活性剂与蛋白质连接的方法。

4. 发明授权

US5049656A Sequential peptide and oligonucleotide syntheses using immunoaffinity techniques 失效
标题翻译：使用免疫亲和技术的顺序肽和寡核苷酸合成
公开(公告)号：US5049656A
公开(公告)日：1991-09-17
申请号：US288009
申请日：1988-12-21
申请人： William Lewis , Jay Stout , Gino Van Heeke , Dwane E. Wylie , Sheldon M. Schuster , Fred W. Wagner
发明人： William Lewis , Jay Stout , Gino Van Heeke , Dwane E. Wylie , Sheldon M. Schuster , Fred W. Wagner
IPC分类号： A61K39/395 , A61K48/00 , C07H21/04 , C07K1/00 , C07K1/02 , C07K1/04 , C07K1/06 , C07K1/113 , C07K1/22 , C07K14/42 , C07K14/575 , C07K14/655 , C07K16/00 , C12N15/20
CPC分类号： C07K1/006 , C07K1/04 , C07K1/22 , Y10S435/803 , Y10S435/81 , Y10S436/824
摘要： The invention is directed to a method for purifying sequentially synthesized peptides and oligonucleotides by immunoaffinity techniques. Selected products are lapped with an antigenic capping agent and are conjugated with antibodies that are specific for the capping agent.
摘要翻译：本发明涉及通过免疫亲和技术纯化顺序合成的肽和寡核苷酸的方法。选择的产物用抗原封端剂研磨，并与针对封端剂特异的抗体缀合。

5. 发明授权

US5221736A Sequential peptide and oligonucleotide syntheses using immunoaffinity techniques 失效
标题翻译：使用免疫亲和技术的顺序肽和寡核苷酸合成
公开(公告)号：US5221736A
公开(公告)日：1993-06-22
申请号：US454372
申请日：1989-12-21
申请人： Thomas R. Coolidge , William Lewis , Sheldon M. Schuster , Dwane Wylie , Fred W. Wagner , Jay Stout , Gino van Heeke
发明人： Thomas R. Coolidge , William Lewis , Sheldon M. Schuster , Dwane Wylie , Fred W. Wagner , Jay Stout , Gino van Heeke
IPC分类号： A61K39/395 , A61K48/00 , C07H21/04 , C07K1/00 , C07K1/02 , C07K1/04 , C07K1/06 , C07K1/113 , C07K1/22 , C07K14/42 , C07K14/575 , C07K14/655 , C07K16/00 , C12N15/20
CPC分类号： C07K1/006 , C07K1/04 , C07K1/22 , Y10S435/803 , Y10S435/81 , Y10S436/824
摘要： The invention is directed to a method of purifying sequentially synthesized peptides and oligonucleotides by affinity techniques. Selected products are capped with and N-terminus capping agent for peptides or a 5'-terminus capping agents for oligonucleotides, and then bound with affinity agents that are selective for the corresponding capping agents.
摘要翻译：本发明涉及通过亲和技术纯化顺序合成的肽和寡核苷酸的方法。所选择的产物与肽封端的N-末端封端剂或寡核苷酸的5'-末端封端剂，然后与对相应的封端剂有选择性的亲和试剂结合。

6. 发明授权

US5464759A Sequential oligonucleotide syntheses using immunoaffinity techniques 失效
标题翻译：使用免疫亲和技术的顺序寡核苷酸合成
公开(公告)号：US5464759A
公开(公告)日：1995-11-07
申请号：US18100
申请日：1993-02-17
申请人： Thomas R. Coolidge , William Lewis , Sheldon M. Schuster , Jay Stout , Gino van Heeke , Dwane Wylie , Fred W. Wagner
发明人： Thomas R. Coolidge , William Lewis , Sheldon M. Schuster , Jay Stout , Gino van Heeke , Dwane Wylie , Fred W. Wagner
IPC分类号： A61K39/395 , A61K48/00 , C07H21/04 , C07K1/00 , C07K1/02 , C07K1/04 , C07K1/06 , C07K1/113 , C07K1/22 , C07K14/42 , C07K14/575 , C07K14/655 , C07K16/00 , C12N15/20 , C12P19/34 , C12Q1/68
CPC分类号： C07K1/006 , C07K1/04 , C07K1/22 , Y10S435/803 , Y10S435/81 , Y10S436/824
摘要： The invention is directed to a method of purifying sequentially synthesized peptides and oligonucleotides by affinity techniques. Selected products are capped with and N-terminus capping agent for peptides or a 5'-terminus capping agents for oligonucleotides, and then bound with affinity agents that are selective for the corresponding capping agents.
摘要翻译：本发明涉及通过亲和技术纯化顺序合成的肽和寡核苷酸的方法。所选择的产物与肽封端的N-末端封端剂或寡核苷酸的5'-末端封端剂，然后与对相应的封端剂有选择性的亲和试剂结合。

7. 发明授权

US5807695A Metallic cation binding polypeptides and methods therefor 失效
标题翻译：金属阳离子结合多肽及其方法
公开(公告)号：US5807695A
公开(公告)日：1998-09-15
申请号：US773688
申请日：1996-12-27
申请人： Fred W. Wagner , Dwane E. Wylie , Sheldon M. Schuster
发明人： Fred W. Wagner , Dwane E. Wylie , Sheldon M. Schuster
IPC分类号： C07K16/44 , G01N33/84 , G01N33/547 , G01N33/531
CPC分类号： C07K16/44 , G01N33/84 , C07K2317/55
摘要： The invention is directed to monoclonal antibodies, their fragments, single chains and polypeptide mimics of their hypervariable regions which immunoreact with bare small moieties such as metallic cations and small organic molecules, the hybridomas for production of the monoclonal antibodies, immunogen compounds for developing the hybridomas, and methods for use of the monoclonal antibodies.
摘要翻译：本发明涉及与裸露小部分如金属阳离子和小有机分子免疫反应的其高变区的单克隆抗体，其片段，单链和多肽模拟物，用于产生单克隆抗体的杂交瘤，用于开发杂交瘤的免疫原化合物以及使用单克隆抗体的方法。

8. 发明授权

US5503987A Monoclonal antibodies specific for small moieties and method for their use 失效
标题翻译：对小部分有特异性的单克隆抗体及其使用方法
公开(公告)号：US5503987A
公开(公告)日：1996-04-02
申请号：US990543
申请日：1992-12-14
申请人： Fred W. Wagner , Dwane E. Wylie , Sheldon M. Schuster
发明人： Fred W. Wagner , Dwane E. Wylie , Sheldon M. Schuster
IPC分类号： C07K14/76 , C07K14/00 , C07K16/00 , C07K16/44 , C12N5/10 , C12N15/02 , C12P21/08 , C12R1/91 , G01N33/53 , G01N33/543 , G01N33/68 , G01N33/84 , G01N33/547 , G01N33/531
CPC分类号： C07K16/44 , G01N33/5308 , G01N33/54353 , G01N33/68 , G01N33/84 , C07K2317/55
摘要： The invention is directed to monoclonal antibodies which immunoreact with bare small moieties such as metallic cations and small organic molecules, the hybridomas for production of the monoclonal antibodies, immunogen compounds for developing the hybridomas, and methods for use of the monoclonal antibodies.
摘要翻译：本发明涉及与裸露小部分例如金属阳离子和小有机分子免疫反应的单克隆抗体，用于产生单克隆抗体的杂交瘤，用于开发杂交瘤的免疫原化合物和用于单克隆抗体的方法。

9. 发明授权

US5639624A Monoclonal antibodies specific for metallic cations and method therefor 失效
标题翻译：金属阳离子特异性单克隆抗体及其方法
公开(公告)号：US5639624A
公开(公告)日：1997-06-17
申请号：US266163
申请日：1994-06-27
申请人： Fred W. Wagner , Dwane E. Wylie , Sheldon M. Schuster
发明人： Fred W. Wagner , Dwane E. Wylie , Sheldon M. Schuster
IPC分类号： C07K16/44 , G01N33/84 , G01N33/53 , C07K16/00 , C12N5/00 , G01N33/549
CPC分类号： C07K16/44 , G01N33/84 , C07K2317/55
摘要： The invention is directed to monoclonal antibodies, their fragments, single chains and polypeptide mimics of their hypervariable regions which immunoreact with bare small moieties such as metallic cations the hybridomas for production of the monoclonal antibodies, immunogen compounds for developing the hybridomas, and methods for use of the monoclonal antibodies.
摘要翻译：本发明涉及它们的高变区的单克隆抗体，其片段，单链和多肽模拟物，其与裸露的小部分例如金属阳离子免疫反应，用于产生单克隆抗体的杂交瘤，用于开发杂交瘤的免疫原化合物和使用方法的单克隆抗体。

10. 发明授权

US5656456A Chemical method for selective modification of the N- and/or C-terminal amino acid .alpha.-carbon reactive group of a recombinant polypeptide or a portion thereof 失效
标题翻译：用于选择性修饰重组多肽或其部分的N-和/或C-末端氨基酸α-碳反应性基团的化学方法
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。

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