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    Process for the production of naturally folded and secreted proteins
    1.
    发明授权
    Process for the production of naturally folded and secreted proteins 失效
    生产天然折叠和分泌蛋白质的方法

    公开(公告)号：US06309861B1

    公开(公告)日：2001-10-30

    申请号：US09553498

    申请日：2000-04-20

    申请人： Dorothee Ambrosius ,  Rainer Rudolph ,  Joerg Schaeffner ,  Elisabeth Schwarz

    发明人： Dorothee Ambrosius ,  Rainer Rudolph ,  Joerg Schaeffner ,  Elisabeth Schwarz

    IPC分类号： C12P2106

    CPC分类号： C12N9/6459 ,  C07K1/1133 ,  C07K2319/00 ,  C07K2319/02 ,  C12N1/20 ,  C12P21/02 ,  C12Y304/21069 ,  Y10S435/849 ,  Y10S435/877

    摘要： A process produces a water-soluble, naturally folded eukaryotic polypeptide containing two or several cysteines linked by disulfide bridges. This process involves culturing prokaryotic cells, a) in which the prokaryotic cells contain an expression vector which encodes the polypeptide which contains a prokaryotic signal sequence at the N-terminus, b) under conditions under which the polypeptide is secreted into the periplasm or the medium, c) cleaving the signal sequence and isolating the polypeptide from the periplasm or the medium. In this process, the culturing is carried out in the presence of arginine or a compound of the formula I R2—CO—NR1 (I) in which R and R1 represent hydrogen or a saturated or unsaturated branched or unbranched C1-C4 alkyl chain and R2 represents hydrogen, NHR1 or a saturated or unsaturated branched or unbranched C1-C3 alkyl chain, is suitable for the recombinant production of polypeptides in prokaryotes in a high yield.

    摘要翻译： 一种方法产生含有两个或几个通过二硫键连接的半胱氨酸的水溶性天然折叠的真核多肽。 该方法包括培养原核细胞，a）其中原核细胞含有编码在N末端含有原核信号序列的多肽的表达载体，b）在多肽分泌到周质或培养基中的条件下 ，c）切割信号序列并从周质或培养基中分离多肽。 在该方法中，在精氨酸或其中R和R 1表示氢或饱和或不饱和的支链或非支链C 1 -C 4烷基链的式I R 2 -CO-NR 1（I）的化合物存在下进行培养， R2代表氢，NHR1或饱和或不饱和的支链或非支链C1-C3烷基链，适合以高产率在原核生物中重组生产多肽。

    Process for the production of naturally folded and secreted proteins by co-secretion of molecular chaperones
    2.
    发明授权
    Process for the production of naturally folded and secreted proteins by co-secretion of molecular chaperones 有权
    通过分子伴侣的共分泌生产天然折叠和分泌的蛋白质的方法

    公开(公告)号：US06455279B1

    公开(公告)日：2002-09-24

    申请号：US09618869

    申请日：2000-07-19

    申请人： Dorothee Ambrosius ,  Rainer Rudolph ,  Joerg Schaeffner ,  Elisabeth Schwarz

    发明人： Dorothee Ambrosius ,  Rainer Rudolph ,  Joerg Schaeffner ,  Elisabeth Schwarz

    IPC分类号： C12P2106

    CPC分类号： C12N9/6459 ,  C07K2319/02 ,  C12N1/20 ,  C12P21/02 ,  C12Y304/21069

    摘要： A process for the production of a naturally folded eukaryotic polypeptide containing two or several cysteines linked by disulfide bridges by a) culturing prokaryotic cells in which the said prokaryotic cells contain an expression vector which codes for the said polypeptide which contains a prokaryotic signal sequence at the N-terminus, b) secreting the polypeptide into the periplasm or the medium, c) cleaving the signal sequence and isolating the polypeptide from the periplasm or the medium, which is characterized in that a nucleic acid coding for a molecular chaperone is additionally expressed in the said prokaryotic cell and the chaperone is secreted into the periplasm, is suitable for the recombinant production of polypeptides in prokaryotes in a high yield.

    摘要翻译： 一种生产包含通过二硫键连接的两个或几个半胱氨酸的天然折叠的真核生物多肽的方法，其方法是：a）培养原核细胞，其中所述原核细胞含有表达载体，所述表达载体编码所述多肽，其含有原核信号序列 N末端，b）将多肽分泌到周质或培养基中，c）切割信号序列并从周质或培养基中分离多肽，其特征在于编码分子伴侣的核酸另外表达于 所述原核细胞和分子伴侣分泌到周质中，适于以高产率重组生产原核生物中的多肽。

    Method for obtaining active pro-NGF and beta-NGF
    3.
    发明授权
    Method for obtaining active pro-NGF and beta-NGF 有权
    获得活性前NGF和β-NGF的方法

    公开(公告)号：US08501439B2

    公开(公告)日：2013-08-06

    申请号：US12765229

    申请日：2010-04-22

    申请人： Anke Rattenholl ,  Adelbert Grossmann ,  Elisabeth Schwarz ,  Rainer Rudolph

    发明人： Anke Rattenholl ,  Adelbert Grossmann ,  Elisabeth Schwarz ,  Rainer Rudolph

    IPC分类号： C12N15/18

    CPC分类号： C07K14/48

    摘要： The invention relates to a method for producing biologically active β-NGF from the proform proNGF. After expressing the proform of the β-NGF in a prokaryotic host cell, the recombinant protein is isolated in the form of insoluble inactive aggregates (inclusion bodies). After the solubilization thereof in a strong denaturing agent and the subsequent conversion thereof into the natural conformation, which is determined by the disulfide bridges present in the natural β-NGF, biologically active β-NGF is obtained by subsequently splitting-off the prosequence.

    摘要翻译： 本发明涉及从前体proNGF生产生物活性β-NGF的方法。 在原核宿主细胞中表达β-NGF的形式后，以不溶性无活性聚集体（包涵体）的形式分离重组蛋白。 在将其溶解在强变性剂中并随后转化成通过存在于天然β-NGF中的二硫键确定的天然构象后，通过随后分离原序列获得生物活性β-NGF。

    Pharmaceutical preparations comprising human proNGF
    4.
    发明授权
    Pharmaceutical preparations comprising human proNGF 有权
    包含人类proNGF的药物制剂

    公开(公告)号：US08318671B1

    公开(公告)日：2012-11-27

    申请号：US09807096

    申请日：1999-10-11

    申请人： Anke Rattenholl ,  Adelbert Grossmann ,  Elisabeth Schwarz ,  Rainer Rudolph

    发明人： Anke Rattenholl ,  Adelbert Grossmann ,  Elisabeth Schwarz ,  Rainer Rudolph

    IPC分类号： C07K14/48 ,  A61K38/18

    CPC分类号： C07K14/48

    摘要： The invention relates to a method for producing biologically active β-NGF from the proform proNGF. After expressing the proform of the β-NGF in a prokaryotic host cell, the recombinant protein is isolated in the form of insoluble inactive aggregates (inclusion bodies). After the solubilization thereof in a strong denaturing agent and the subsequent conversion thereof into the natural conformation, which is determined by the disulfide bridges present in the natural β-NGF, biologically active β-NGF is obtained by subsequently splitting-off the prosequence.

    摘要翻译： 本发明涉及从前体proNGF生产生物活性的NGF的方法。 在原核宿主细胞中表达生物素-NGF的形式后，重组蛋白以不溶性非活性聚集体（包涵体）的形式分离。 在将其溶解在强变性剂中并随后转化成通过存在于天然和/或生长因子中的二硫键确定的天然构象后，通过随后分离原序列获得生物活性 。

    METHOD FOR OBTAINING ACTIVE BETA-NGF
    5.
    发明申请
    METHOD FOR OBTAINING ACTIVE BETA-NGF 有权
    获得活性β-NGF的方法

    公开(公告)号：US20100203589A1

    公开(公告)日：2010-08-12

    申请号：US12765229

    申请日：2010-04-22

    申请人： Anke Rattenholl ,  Adelbert Grossmann ,  Elisabeth Schwarz ,  Rainer Rudolph

    发明人： Anke Rattenholl ,  Adelbert Grossmann ,  Elisabeth Schwarz ,  Rainer Rudolph

    IPC分类号： C12P21/02

    CPC分类号： C07K14/48

    摘要： The invention relates to a method for producing biologically active β-NGF from the proform proNGF. After expressing the proform of the β-NGF in a prokaryotic host cell, the recombinant protein is isolated in the form of insoluble inactive aggregates (inclusion bodies). After the solubilization thereof in a strong denaturing agent and the subsequent conversion thereof into the natural conformation, which is determined by the disulfide bridges present in the natural β-NGF, biologically active β-NGF is obtained by subsequently splitting-off the prosequence.

    摘要翻译： 本发明涉及从前体proNGF生产生物活性的NGF的方法。 在原核宿主细胞中表达生物素-NGF的形式后，重组蛋白以不溶性非活性聚集体（包涵体）的形式分离。 在将其溶解在强变性剂中并随后转化成通过存在于天然和/或生长因子中的二硫键确定的天然构象后，通过随后分离前体 。

    Production of recombinant BMP-2
    6.
    发明授权
    Production of recombinant BMP-2 失效
    重组BMP-2的生产

    公开(公告)号：US07354901B2

    公开(公告)日：2008-04-08

    申请号：US10433077

    申请日：2001-11-27

    申请人： Rainer Rudolph ,  Elisabeth Schwarz ,  Gerhard Herr ,  Frank Hillger

    发明人： Rainer Rudolph ,  Elisabeth Schwarz ,  Gerhard Herr ,  Frank Hillger

    IPC分类号： C07K14/51 ,  A61K38/00 ,  A61K35/32

    CPC分类号： C07K14/51

    摘要： In order to recombinantly prepare a biologically active protein of the TGF-β superfamily, a protein, whose amino terminus consists of the pro sequence of a protein of the TGF-β superfamily, or parts thereof, to which the mature domain of this protein or of another protein of TGF-β superfamily which exhibits at least 35% homology with mature BMP-2 is attached, is expressed in prokaryotes under conditions in which at least a part of the protein is obtained in the form of inclusion bodies, the inclusion bodies are isolated and solubilized under denaturing conditions, the denatured, monomeric and biologically inactive protein which has been solubilized from the inclusion bodies is renatured, with folding and dimerization to give the soluble, biologically active conformation and, where appropriate, after the renaturation, the mature protein is released proteolytically from its pro form.

    摘要翻译： 为了重组制备TGF-β超家族的生物活性蛋白，一种蛋白质，其氨基末端由TGF-β超家族的蛋白质的前序列或其部分组成，其中该蛋白质的成熟结构域或 与成熟BMP-2呈现至少35％同源性的另一种TGF-β超家族的蛋白质被连接，在原核生物中在以包涵体形式获得至少一部分蛋白质的条件下表达，包涵体 在变性条件下分离和溶解，已经从包涵体溶解的变性，单体和生物活性蛋白质被复性，具有折叠和二聚作用以产生可溶性的生物活性构象，并且在适当的情况下在复性后成熟 蛋白质从其形式蛋白水解释放。

    Recombinant core-streptavidin
    7.
    发明授权
    Recombinant core-streptavidin 失效
    重组核心 - 链霉抗生物素蛋白

    公开(公告)号：US5489528A

    公开(公告)日：1996-02-06

    申请号：US211833

    申请日：1994-04-28

    申请人： Erhard Kopetzki ,  Rainer Rudolph ,  Adelbert Grossmann

    发明人： Erhard Kopetzki ,  Rainer Rudolph ,  Adelbert Grossmann

    IPC分类号： C07K14/195 ,  C07K14/36 ,  C07K14/41 ,  C12N1/21 ,  C12N15/09 ,  C12N15/31 ,  C12P21/02 ,  C12R1/19 ,  G01N33/50 ,  G01N33/53 ,  C12N5/10 ,  C12N15/63

    CPC分类号： C07K14/36

    摘要： The present invention concerns a process for the isolation of recombinant core streptavidin in which host cells are transformed with a DNA coding for core streptavidin, the transformed host cells are cultured under suitable conditions, the DNA coding for core streptavidin is expressed and the recombinant core streptavidin is isolated from the host cells or the culture medium, wherein a DNA coding for core streptavidin is used which has(a) the nucleotide sequence shown in SEQ ID NO. 1 or(b) a nucleotide sequence corresponding to the nucleotide sequence (a) within the scope of the degeneracy of the genetic code.

    摘要翻译： PCT No.PCT / EP92 / 02463。 371日期1994年04月28日 102（e）日期1994年4月28日PCT提交1992年10月28日PCT公布。 公开号WO93 / 09144 日本1993年5月13日。本发明涉及分离重组核心链亲和素的方法，其中宿主细胞用编码核心链霉亲和素的DNA转化，转化的宿主细胞在合适的条件下培养，编码核心链霉抗生物素蛋白的DNA为 表达，并且从宿主细胞或培养基中分离重组核心链霉抗生物素蛋白，其中使用编码核心链亲和素的DNA，其具有（a）SEQ ID NO： 1或（b）对应于遗传密码简并范围内的核苷酸序列（a）的核苷酸序列。