公开(公告)号：US20020081735A1

公开(公告)日：2002-06-27

申请号：US09899999

申请日：2001-07-09

申请人： BioNebraska, Inc.

发明人： Yuannan Xia

IPC分类号： C12N005/06 ,  C12N015/00 ,  C12N009/16 ,  C07H021/04

CPC分类号： C07K14/005 ,  C07K14/605 ,  C12N15/70 ,  C12N15/8216 ,  C12N2710/00022

摘要： The present invention provides novel promoter sequences obtained from Chlorella virus. The invention includes gene constructs comprising a promoter sequence of the invention operably linked to a DNA sequence encoding a structural gene. The invention also provides vectors and host cells for expressing product encoded by the structural gene of a gene construct of the invention amd cells transformed with the heterologous gene operably linked to the promoter.

摘要翻译： 本发明提供从小球藻病毒获得的新型启动子序列。 本发明包括可操作地连接到编码结构基因的DNA序列的本发明的启动子序列的基因构建体。 本发明还提供用于表达由本发明的基因构建体的结构基因编码的产物的载体和宿主细胞，所述细胞用可操作地连接于启动子的异源基因转化。

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：US5972656A

公开(公告)日：1999-10-26

申请号：US888366

申请日：1997-07-03

申请人： Osvaldo Lopez ,  Dwane E. Wylie ,  Fred W. Wagner

发明人： Osvaldo Lopez ,  Dwane E. Wylie ,  Fred W. Wagner

IPC分类号： A62D3/00 ,  A62D3/33 ,  A62D101/24 ,  A62D101/43 ,  G01N33/53 ,  C07K16/00 ,  C07K16/44 ,  C12P21/04

CPC分类号： A62D3/33 ,  A62D2101/24 ,  A62D2101/43 ,  C07K2317/55

摘要： Metal binding polypeptides which include an amino acid sequence coding for a variable region of a monoclonal antibody which immunoreacts with a mercury cation and nucleotides which include a nucleic acid sequence coding for the variable region are provided. The invention is also directed to fusion proteins which include a phage coat protein or portion thereof and the monoclonal antibody heavy chain variable region. The invention also provides bacteriophages which include the fusion protein in their coat. In addition, methods for detecting, removing, adding, or neutralizing mercuric cations in biological or inanimate systems through the use of the mercury binding polypeptides are provided.

摘要翻译： 提供了包括编码与汞阳离子免疫反应的单克隆抗体的可变区的氨基酸序列的氨基酸序列和包含编码可变区的核酸序列的核苷酸的金属结合多肽。 本发明还涉及包含噬菌体外壳蛋白或其部分和单克隆抗体重链可变区的融合蛋白。 本发明还提供了在其外壳中包含融合蛋白的噬菌体。 此外，提供了通过使用汞结合多肽检测，除去，添加或中和生物或无生物体系中的汞阳离子的方法。

公开(公告)号：US5512459A

公开(公告)日：1996-04-30

申请号：US95162

申请日：1993-07-20

申请人： Fred W. Wagner ,  Jay Stout ,  Dennis Henriksen ,  Bruce Partridge ,  Shane Manning

发明人： Fred W. Wagner ,  Jay Stout ,  Dennis Henriksen ,  Bruce Partridge ,  Shane Manning

IPC分类号： C12N15/09 ,  A61K38/00 ,  A61K38/04 ,  A61P3/08 ,  A61P13/02 ,  A61P15/00 ,  A61P43/00 ,  C07K14/60 ,  C07K14/605 ,  C12N1/21 ,  C12P21/02 ,  C12P21/06 ,  C12R1/19 ,  C12N9/74 ,  C12N9/76

CPC分类号： C07K14/60 ,  C07K14/605 ,  C12P21/02 ,  C12P21/06 ,  A61K38/00 ,  C07K2319/00

摘要： The method of the invention provides for the formation of a recombinant polypeptide which has been modified at the C-terminal end through the use of a transpeptidation process. The method is suitable for modifying recombinant polypeptides of any source including those which may be commercially available, those derived from recombinant single copy or multicopy polypeptide constructs, or those derived from single or multicopy recombinant fusion protein constructs. The transpeptidation reaction involves contacting an endopeptidase enzyme with a recombinant polypeptide to substitute an addition unit, of one or more amino acids, for a leaving unit, linked to a core polypeptide through a cleavage site recognized by the endopeptidase enzyme. Recombinant polypeptides derived from multicopy polypeptide constructs may be cleaved from the multicopy polypeptide at the N-terminal and C-terminal ends and simultaneously under go substitution of the leaving unit by the desired addition unit. The invention utilizes known and newly discovered cleavage recognition sites to effectuate the desired modification products.

摘要翻译： 本发明的方法提供了通过使用转肽酶方法在C末端修饰的重组多肽的形成。 该方法适用于修饰任何来源的重组多肽，包括可商购的重组多肽，衍生自重组单拷贝或多拷贝多肽构建体的重组多肽，或衍生自单拷贝或多拷贝重组融合蛋白构建体的重组多肽。 转肽反应包括使内肽酶与重组多肽接触，以将一个或多个氨基酸的加成单元替换为通过内肽酶识别的切割位点与核心多肽连接的离去单元。 衍生自多拷贝多肽构建体的重组多肽可以在N末端和C-末端的多拷贝多肽中切割，同时在离去单位取代所需的加成单位。 本发明利用已知和新发现的裂解识别位点来实现所需的修饰产物。

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：US6037143A

公开(公告)日：2000-03-14

申请号：US967374

申请日：1997-11-07

申请人： Fred W. Wagner ,  Jay Stout ,  Dennis Henriksen ,  Bruce Partridge ,  Shane Manning

发明人： Fred W. Wagner ,  Jay Stout ,  Dennis Henriksen ,  Bruce Partridge ,  Shane Manning

IPC分类号： C12N15/09 ,  A61K38/00 ,  A61K38/04 ,  A61P3/08 ,  A61P13/02 ,  A61P15/00 ,  A61P43/00 ,  C07K14/60 ,  C07K14/605 ,  C12N1/21 ,  C12P21/02 ,  C12P21/06 ,  C12R1/19

CPC分类号： C07K14/60 ,  C07K14/605 ,  C12P21/02 ,  C12P21/06 ,  A61K38/00 ,  C07K2319/00

摘要： The method of the invention provides for the formation of a recombinant polypeptide which has been modified at the C-terminal end through the use of a transpeptidation process. The method is suitable for modifying recombinant polypeptides of any source including those which may be commercially available, those derived from recombinant single copy or multicopy polypeptide constructs, or those derived from single or multicopy recombinant fusion protein constructs. The transpeptidation reaction involves contacting an endopeptidase enzyme with a recombinant polypeptide to substitute an addition unit, of one or more amino acids, for a leaving unit, linked to a core polypeptide through a cleavage site recognized by the endopeptidase enzyme. Recombinant polypeptides derived from multicopy polypeptide constructs may be cleaved from the multicopy polypeptide at the N-terminal and C-terminal ends and simultaneously under go substitution of the leaving unit by the desired addition unit. The invention utilizes known and newly discovered cleavage recognition sites to effectuate the desired modification products.

公开(公告)号：US5595887A

公开(公告)日：1997-01-21

申请号：US552810

申请日：1990-07-16

申请人： Thomas R. Coolidge ,  Fred Wagner ,  Gino van Heeke ,  Sheldon M. Schuster ,  Jay Stout ,  Dwane E. Wylie

发明人： Thomas R. Coolidge ,  Fred Wagner ,  Gino van Heeke ,  Sheldon M. Schuster ,  Jay Stout ,  Dwane E. Wylie

IPC分类号： C07K14/575 ,  C07K1/22 ,  C07K7/14 ,  C07K14/00 ,  C07K14/245 ,  C07K14/47 ,  C07K14/585 ,  C07K14/595 ,  C07K19/00 ,  C12N1/21 ,  C12N9/14 ,  C12N9/88 ,  C12N15/09 ,  C12N15/62 ,  C12P21/00 ,  C12R1/19 ,  C12P21/04 ,  C12P21/06

CPC分类号： C12N9/14 ,  C07K14/245 ,  C07K14/47 ,  C07K14/585 ,  C07K14/595 ,  C07K7/14 ,  C12N15/62 ,  C12N9/88 ,  C07K2319/00 ,  C07K2319/50 ,  C07K2319/75

摘要： Methods are presented for producing and purifying a variable fusion polypeptide which can be purified by affinity chromatography with the binding protein partner. The variable fusion polypeptide construct has tandem coupled segments containing one or more copies of a desired peptide linked to carbonic anhydrase as the purification binding protein. In the methods, the fusion protein is expressed in a recombinant host using a recombinant vector containing a gene encoding the fusion polypeptide. Then the expressed fusion polypeptide is purified by immobilized reversible inhibitor affinity chromatography. Finally, the purified fusion polypeptide is cleaved from the desired peptides by chemical or enzymatic means and the desired peptides purified with affinity chromatography.

摘要翻译： 提供了用于产生和纯化可通过用结合蛋白配偶体进行亲和层析纯化的可变融合多肽的方法。 可变融合多肽构建体具有串联耦合区段，其含有与碳酸酐酶连接的期望肽的一个或多个拷贝作为纯化结合蛋白。 在该方法中，使用含有编码融合多肽的基因的重组载体，在重组宿主中表达融合蛋白。 然后通过固定的可逆抑制剂亲和层析纯化表达的融合多肽。 最后，通过化学或酶法将纯化的融合多肽从期望的肽切割，并用亲和层析纯化所需的肽。

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：US6111079A

公开(公告)日：2000-08-29

申请号：US767128

申请日：1996-12-04

申请人： Dwane E. Wylie ,  Osvaldo Lopez ,  Peter Joseph Murray ,  Peter Goebel

发明人： Dwane E. Wylie ,  Osvaldo Lopez ,  Peter Joseph Murray ,  Peter Goebel

IPC分类号： C07K16/44 ,  C12P21/08 ,  C07H21/02 ,  C07H21/04

CPC分类号： C07K16/44 ,  C07K2319/00

摘要： Metal binding polypeptides which include an amino acid sequence coding for a light chain variable region of a monoclonal antibody capable of immunoreacting with a lead cation and nucleotides which include a nucleic acid sequence coding for the variable region are provided. The invention is also directed to fusion proteins and Fab fragments which include the light chain variable region.

摘要翻译： 提供了包含编码能够与铅阳离子进行免疫反应的单克隆抗体的轻链可变区的氨基酸序列的金属结合多肽和包含编码可变区的核酸序列的核苷酸。 本发明还涉及包含轻链可变区的融合蛋白和Fab片段。

公开(公告)号：US5846774A

公开(公告)日：1998-12-08

申请号：US821559

申请日：1997-03-21

申请人： Yuannan Xia

发明人： Yuannan Xia

IPC分类号： A01H5/00 ,  C07K14/01 ,  C07K14/605 ,  C12N1/21 ,  C12N5/10 ,  C12N15/09 ,  C12N15/70 ,  C12N15/82 ,  C12P21/06 ,  C12N1/00 ,  C12N15/11

CPC分类号： C07K14/005 ,  C07K14/605 ,  C12N15/70 ,  C12N15/8216 ,  C12N2770/24322

摘要： The present invention provides novel promoter sequences obtained from Chlorella virus. The invention includes gene constructs comprising a promoter sequence of the invention operably linked to a DNA sequence encoding a structural gene. The invention also provides vectors and host cells for expressing a product encoded by the structural gene of a gene construct of the invention and cells transformed with the heterologous gene operably linked to the promoter.

摘要翻译： 本发明提供从小球藻病毒获得的新型启动子序列。 本发明包括可操作地连接到编码结构基因的DNA序列的本发明的启动子序列的基因构建体。 本发明还提供用于表达由本发明的基因构建体的结构基因编码的产物的载体和宿主细胞，以及用与启动子可操作连接的异源基因转化的细胞。