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    Synthesis of protein with an identification peptide, and hybrid polypeptide incorporating same
    1.
    发明授权
    Synthesis of protein with an identification peptide, and hybrid polypeptide incorporating same 失效
    用鉴定肽合成蛋白质，以及掺入其的杂交多肽

    公开(公告)号：US4782137A

    公开(公告)日：1988-11-01

    申请号：US76811

    申请日：1987-07-23

    申请人： Thomas P. Hopp ,  Susan L. Bektesh ,  Paul J. Conlon, III ,  Carl J. March

    发明人： Thomas P. Hopp ,  Susan L. Bektesh ,  Paul J. Conlon, III ,  Carl J. March

    IPC分类号： C07K14/55 ,  C07K16/44 ,  C12N15/62 ,  A61K37/02 ,  C12P21/00

    CPC分类号： C07K14/55 ,  C07K16/44 ,  C12N15/62 ,  C07K2319/00 ,  C07K2319/40 ,  C07K2319/50

    摘要： A hybrid polypeptide composed of an identification peptide and a desired functional protein are produced by recombinant DNA techniques. A DNA expression vector is constructed that includes segments of DNA coding for the identification peptide and the desired functional protein. The identification peptide consists of a highly antigenic N-terminal portion and a C-terminal linking portion that connects the identification peptide to the N-terminal of the functional protein. The linking portion of the identification peptide is cleavable at a specific amino acid residue adjacent the functional protein by use of a sequence specific proteolytic enzyme or chemical proteolytic agent. The hybrid polypeptide expressed by the host cells transformed by the cloning vector is removed therefrom and purfied by affinity chromatography techniques by use of an immobilized ligand specific to the antigenic portion of the identification peptide. The protein is then cleaved from the isolated hybrid polypeptide with an appropriate proteolic enzyme or chemical agent, thereby releasing the mature functional protein in highly purified, highly active state.

    摘要翻译： 通过重组DNA技术产生由鉴定肽和所需功能性蛋白质组成的杂合多肽。 构建DNA表达载体，其包括编码鉴定肽和所需功能性蛋白质的DNA片段。 识别肽由高度抗原的N末端部分和连接识别肽与功能性蛋白质的N末端的C-末端连接部分组成。 识别肽的连接部分可通过使用序列特异性蛋白水解酶或化学蛋白水解剂在与功能蛋白相邻的特定氨基酸残基处可切割。 由克隆载体转化的宿主细胞表达的杂合多肽从其中除去，并通过使用对鉴定肽的抗原部分特异性的固定化配体的亲和层析技术进行纯化。 然后用合适的蛋白酶或化学试剂从分离的杂交多肽切割蛋白质，从而释放高度纯化，高活性状态的成熟功能性蛋白质。

    Synthesis of protein with an identification peptide
    2.
    发明授权
    Synthesis of protein with an identification peptide 失效
    用鉴定肽合成蛋白质

    公开(公告)号：US4703004A

    公开(公告)日：1987-10-27

    申请号：US573825

    申请日：1984-01-24

    申请人： Thomas P. Hopp ,  Susan L. Bektesh ,  Paul J. Conlon, III ,  Carl J. March

    发明人： Thomas P. Hopp ,  Susan L. Bektesh ,  Paul J. Conlon, III ,  Carl J. March

    IPC分类号： G01N33/531 ,  A61K35/74 ,  A61K36/06 ,  A61K38/00 ,  A61K39/00 ,  C07C15/04 ,  C07H21/04 ,  C07K14/005 ,  C07K14/195 ,  C07K14/55 ,  C07K16/00 ,  C07K16/44 ,  C07K19/00 ,  C12N5/00 ,  C12N5/10 ,  C12N15/00 ,  C12N15/09 ,  C12N15/62 ,  C12P21/00 ,  C12P21/02 ,  C12P21/08 ,  C12R1/19 ,  C12R1/865 ,  C12R1/91 ,  G01N33/577 ,  C12N1/00

    CPC分类号： C07K14/55 ,  C07K16/44 ,  C12N15/62 ,  C07K2319/00 ,  C07K2319/40 ,  C07K2319/50 ,  Y10S930/28

    摘要： A hybrid polypeptide composed of an identification peptide and a desired functional protein are produced by recombinant DNA techniques. A DNA expression vector is constructed that includes segments of DNA coding for the identification peptide and the desired functional protein. The identification peptide consists of a highly antigenic N-terminal portion and a C-terminal linking portion that connects the identification peptide to the N-terminal of the functional protein. The linking portion of the identification peptide is cleavable at a specific amino acid residue adjacent the functional protein by use of a sequence specific proteolytic enzyme or chemical proteolytic agent. The hybrid polypeptide expressed by the host cells transformed by the cloning vector is removed therefrom and purified by affinity chromatography techniques by use of an immobilized ligand specific to the antigenic portion of the identification peptide. The protein is then cleaved from the isolated hybrid polypeptide with an appropriate proteolic enzyme or chemical agent, thereby releasing the mature functional protein in highly purified, highly active state.

    摘要翻译： 通过重组DNA技术产生由鉴定肽和所需功能性蛋白质组成的杂合多肽。 构建DNA表达载体，其包括编码鉴定肽和所需功能性蛋白质的DNA片段。 识别肽由高度抗原的N末端部分和连接识别肽与功能性蛋白质的N末端的C-末端连接部分组成。 识别肽的连接部分可通过使用序列特异性蛋白水解酶或化学蛋白水解剂在与功能蛋白相邻的特定氨基酸残基处可切割。 由克隆载体转化的宿主细胞表达的杂合多肽从其中除去，并通过亲和层析技术通过使用对鉴定肽的抗原部分特异性的固定化配体进行纯化。 然后用合适的蛋白酶或化学试剂从分离的杂交多肽切割蛋白质，从而释放高度纯化，高活性状态的成熟功能性蛋白质。

    Gene encoding biologically active human interleukin 1
    3.
    发明授权
    Gene encoding biologically active human interleukin 1 失效
    基因编码生物活性人白细胞介素1

    公开(公告)号：US5122459A

    公开(公告)日：1992-06-16

    申请号：US443399

    申请日：1989-11-30

    申请人： Paul J. Conlon, III ,  David j. Cosman ,  Kenneth H. Grabstein ,  Thomas P. Hopp ,  Shirley R. Kronheim ,  Alf D. Larsen ,  Carl J. March ,  Virginia L. Price ,  Douglas P. Cerretti

    发明人： Paul J. Conlon, III ,  David j. Cosman ,  Kenneth H. Grabstein ,  Thomas P. Hopp ,  Shirley R. Kronheim ,  Alf D. Larsen ,  Carl J. March ,  Virginia L. Price ,  Douglas P. Cerretti

    IPC分类号： C07K14/545 ,  C12N15/25 ,  C12N15/81

    CPC分类号： C07K14/545 ,  C12N15/81 ,  Y10S435/942

    摘要： Double-stranded cDNA is prepared from polyadenylated RNA extracted from activated human peripheral blood adherent mononuclear cells. The cDNA is inserted within a plasmid vector and then the recombinant plasmid employed to transform an appropriate host. Transformed hosts are identified and grouped into pools. Plasmid DNA prepared from these pools is hybridized with a labeled, synthetic oligonucleotide probe corresponding to a portion of the amino acid sequence of the interleukin 1 protein. Pools of host cells that provide a positive signal to the probe are identified, plated out and then employed in direct bacterial colony hybridization with the same probe, thereby to isolate the particular positive colony. Plasmid DNA is prepared from this colony and characterized by restriction enzyme mapping and sequencing by chain-termination method. The coding region for the IL-1 gene is inserted into a shuttle vector for amplification of the vector followed by expression of functional IL-1.

    摘要翻译： 双链cDNA由从活化的人外周血粘附单核细胞提取的聚腺苷酸化RNA制备。 将cDNA插入质粒载体中，然后将重组质粒用于转化合适的宿主。 已识别转化的主机并将其分组到池中。 从这些池制备的质粒DNA与对应于白介素1蛋白的氨基酸序列的一部分的标记的合成的寡核苷酸探针杂交。 确定提供探针的正信号的宿主细胞池，电镀，然后用于与相同探针的直接细菌菌落杂交，从而分离出特定的阳性菌落。 从该菌落制备质粒DNA，并通过限制酶作图和通过链终止法进行测序。 将用于IL-1基因的编码区插入穿梭载体中用于扩增载体，随后表达功能性IL-1。

    Process for producing homogeneous colony stimulating factor
    5.
    发明授权
    Process for producing homogeneous colony stimulating factor 失效
    生产均质殖民地刺激因子的方法

    公开(公告)号：US4658018A

    公开(公告)日：1987-04-14

    申请号：US588940

    申请日：1984-03-13

    申请人： David L. Urdal ,  Carl J. March ,  Diane Y. Mochizuki ,  Paul J. Conlon, III

    发明人： David L. Urdal ,  Carl J. March ,  Diane Y. Mochizuki ,  Paul J. Conlon, III

    IPC分类号： C07K14/53 ,  C07K3/20 ,  C12P21/00

    CPC分类号： C07K14/53 ,  Y10S930/145

    摘要： Colony stimulating factor derived from malignant cells has been purified by use of various techniques including multiple high performance liquid chromotography steps. By this technique, colony stimulating factor has been resolved into distinct species, and one of the species denominated as CSF-2A, has been purified to homogeneity. The high purification of the CSF-2A has enabled the amino acid composition of this protein molecule to be partially sequenced with an automated sequencer.

    摘要翻译： 来自恶性细胞的集落刺激因子已经通过使用包括多个高效液相色谱步骤的各种技术进行了纯化。 通过这种技术，集落刺激因子已经被分解成不同的物种，并且被称为CSF-2A的物种之一已经被纯化到同质性。 CSF-2A的高纯化使得该蛋白质分子的氨基酸组成能够用自动测序仪部分测序。

    TNF-&agr; converting enzyme
    6.
    发明授权
    TNF-&agr; converting enzyme 失效
    TNF-α转化酶

    公开(公告)号：US06406877B2

    公开(公告)日：2002-06-18

    申请号：US09727169

    申请日：2000-11-29

    申请人： Roy A. Black ,  Charles Rauch ,  Carl J. March ,  Douglas P. Cerretti

    发明人： Roy A. Black ,  Charles Rauch ,  Carl J. March ,  Douglas P. Cerretti

    IPC分类号： C12Q137

    CPC分类号： C12N9/6489 ,  C07K16/40

    摘要： A metalloprotease that converts TNF-&agr; from the 26 kD cell form to the 17 kD form has been isolated and purified and the cDNA sequence known. In particular, the protease has a molecular weight of approximately 80 kD. The isolated and purified protease is useful for designing an inhibitor thereof, and may find use as a therapeutic agent. Assays for detecting the protease-inhibiting activity of a molecule are also an aspect of the invention.

    摘要翻译： 已经分离和纯化将来自26kD细胞形式的TNF-α转化为17kD形式的金属蛋白酶，并且已知cDNA序列。 特别地，蛋白酶的分子量约为80kD。 分离和纯化的蛋白酶可用于设计其抑制剂，可用作治疗剂。 用于检测分子的蛋白酶抑制活性的测定法也是本发明的一个方面。

    Interleukin 2 receptor
    10.
    发明授权
    Interleukin 2 receptor 失效
    白细胞介素2受体

    公开(公告)号：US4578335A

    公开(公告)日：1986-03-25

    申请号：US612559

    申请日：1984-05-21

    申请人： David L. Urdal ,  Carl J. March ,  Steven K. Dower

    发明人： David L. Urdal ,  Carl J. March ,  Steven K. Dower

    IPC分类号： C12N15/09 ,  C07K1/20 ,  C07K1/22 ,  C07K14/00 ,  C07K14/715 ,  C07K16/28 ,  C12P21/00 ,  C12P21/02 ,  C12R1/91 ,  C07K3/02

    CPC分类号： C07K14/7155 ,  C07K16/2866 ,  C07K2317/73 ,  Y10S435/948

    摘要： Interleukin-2 receptor derived from normal and malignant cells has been purified by use of various techniques including affinity chromotography in conjunction with a monoclonal antibody directed to the receptor. Purification also involved reversed phased high performance liquid chromotography. By these techniques, interleukin-2 receptor has been purified to homogeneity. The high purification of the interleukin-2 receptor has enabled this protein molecule to be partially sequenced.

    摘要翻译： 来自正常和恶性细胞的白细胞介素-2受体已经通过使用各种技术进行纯化，包括亲和色谱法与针对受体的单克隆抗体结合。 纯化也涉及反相高效液相色谱法。 通过这些技术，白细胞介素-2受体已经被纯化到均一性。 白细胞介素-2受体的高纯化使得该蛋白质分子被部分测序。