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    Method for producing biologically active human factor VIII in the milk of transgenic animals driven by mammary-specific expression cassettes
    2.
    发明申请
    Method for producing biologically active human factor VIII in the milk of transgenic animals driven by mammary-specific expression cassettes 有权
    在乳腺特异性表达盒驱动的转基因动物的乳中生产生物活性人因子VIII的方法

    公开(公告)号：US20050229261A1

    公开(公告)日：2005-10-13

    申请号：US10820777

    申请日：2004-04-09

    申请人： Winston Cheng ,  Chuan-Mu Chen ,  Shwu-Wha Lin ,  Chih-Hong Wang ,  Chih-Jen Lin ,  Shinn-Chih Wu

    发明人： Winston Cheng ,  Chuan-Mu Chen ,  Shwu-Wha Lin ,  Chih-Hong Wang ,  Chih-Jen Lin ,  Shinn-Chih Wu

    IPC分类号： A01K67/027 ,  C07K14/755 ,  C12N15/85

    CPC分类号： C12N15/8509 ,  A01K67/0275 ,  A01K2217/05 ,  A01K2267/01 ,  C07K14/755 ,  C12N2830/008

    摘要： Hemophilia A is one of the major inherited bleeding disorders caused by a deficiency or abnormality in coagulation factor VIII (FVIII). Hemophiliacs have been treated with whole plasma or purified FVIII concentrates. The risk of transmitting blood-borne viruses and the cost of highly purified FVIII are the major factors that restrict prophylaxis in hemophilia therapy. One of the challenges created by the biotechnology revolution is the development of methods for the economical production of highly purified proteins in large scales. The present invention provides improved mammary expression cassettes useful for the expression of genes at high levels in the milk of transgenic animals. In particular, the present invention provides recombinant signal peptide sequences derived from a-lactalbumin and aS1-casein milk genes suitable for leading protein secretion in the mammary gland. These gene cassettes are capable of delivering different transgenic constructs which result in the production of full-length or B domain-deleted therapeutic levels of biologically active human FVIII in the transgenic animals in vivo. Within the scope of the invention are also method for producing the transgenic non-human mammal, such as mouse, rat, rabbit, goat, sheep, pig and bovine species, capable of expressing human FVIII, and methods of making milk and methods of identifying protein from the transgenic milk.

    摘要翻译： 血友病A是凝血因子VIII（FVIII）缺乏或异常引起的主要遗传性出血性疾病之一。 已经用全血浆或纯化的FVIII浓缩物治疗血友病患者。 传播血源性病毒的风险和高度纯化的FVIII的成本是限制血友病治疗预防的主要因素。 生物技术革命带来的挑战之一是开发大规模经济生产高纯度蛋白质的方法。 本发明提供了可用于在转基因动物的牛奶中高水平表达基因的改进的乳腺表达盒。 特别地，本发明提供了衍生自α-乳白蛋白的重组信号肽序列和适合于在乳腺中引导蛋白质分泌的α1酪蛋白乳基因。 这些基因盒能够递送不同的转基因构建体，其导致在转基因动物体内产生全长或B结构域缺失的生物活性人类FVIII的治疗水平。 在本发明的范围内，也是用于生产能够表达人FVIII的转基因非人哺乳动物如小鼠，大鼠，兔，山羊，绵羊，猪和牛物种的方法，以及制备牛奶的方法和鉴定方法 来自转基因乳的蛋白质。

    Method for expressing multiple recombinant proteins in milk of transgenic non-human mammals
    5.
    发明申请
    Method for expressing multiple recombinant proteins in milk of transgenic non-human mammals 有权
    用于在转基因非人哺乳动物的乳中表达多种重组蛋白的方法

    公开(公告)号：US20050114911A1

    公开(公告)日：2005-05-26

    申请号：US10790720

    申请日：2004-03-03

    申请人： Shinn-Chih Wu ,  Teng-Kuei Cheng ,  Chuan-Mu Chen ,  Shau-Ping Lin ,  Chon-Ho Yen ,  Ping-Cheng Yang

    发明人： Shinn-Chih Wu ,  Teng-Kuei Cheng ,  Chuan-Mu Chen ,  Shau-Ping Lin ,  Chon-Ho Yen ,  Ping-Cheng Yang

    IPC分类号： A01K67/027

    CPC分类号： A01K67/0275 ,  A01K2217/05 ,  A01K2227/108 ,  A01K2267/01

    摘要： The present invention discloses a method for expressing multiple recombinant proteins in transgenic non-human mammalian milk, characterized in which human clotting factor IX gene and porcine lactoferrin gene are transferred into the mammal by gene injection and embryonic implantation to obtain expression in the milk of transgenic mammal and its filial generation. The method of this invention can maintain the stable expression of multiple recombinant proteins in the transgenic mammal during lactation and stable expression amount proximate to that of the first generation in the offsprings of the transgenic non-human mammal.

    摘要翻译： 本发明公开了一种在转基因非人哺乳动物乳中表达多种重组蛋白的方法，其特征在于通过基因注射和胚胎移植将人凝血因子IX基因和猪乳铁蛋白基因转移到哺乳动物中，以获得转基因乳中的表达 哺乳动物及其子代。 本发明的方法可以在哺乳期间保持转基因哺乳动物中多种重组蛋白的稳定表达，并且在转基因非人哺乳动物的后代中可以保持接近于第一代的稳定表达量。

    Transgenic mammal secreting B-domain deleted human FVII in its milk
    7.
    发明授权
    Transgenic mammal secreting B-domain deleted human FVII in its milk 有权
    转基因哺乳动物在其牛奶中分泌B结构域缺失的人FVII

    公开(公告)号：US07667089B2

    公开(公告)日：2010-02-23

    申请号：US10820777

    申请日：2004-04-09

    申请人： Winston T. K. Cheng ,  Chuan-Mu Chen ,  Shwu-Wha Lin ,  Chih-Hong Wang ,  Chih-Jen Lin ,  Shinn-Chih Wu

    发明人： Winston T. K. Cheng ,  Chuan-Mu Chen ,  Shwu-Wha Lin ,  Chih-Hong Wang ,  Chih-Jen Lin ,  Shinn-Chih Wu

    IPC分类号： C12P21/00 ,  A01K67/027 ,  C12N15/00

    CPC分类号： C12N15/8509 ,  A01K67/0275 ,  A01K2217/05 ,  A01K2267/01 ,  C07K14/755 ,  C12N2830/008

    摘要： Hemophilia A is one of the major inherited bleeding disorders caused by a deficiency or abnormality in coagulation factor VIII (FVIII). Hemophiliacs have been treated with whole plasma or purified FVIII concentrates. The risk of transmitting blood-borne viruses and the cost of highly purified FVIII are the major factors that restrict prophylaxis in hemophilia therapy. One of the challenges created by the biotechnology revolution is the development of methods for the economical production of highly purified proteins in large scales. The present invention provides improved mammary expression cassettes useful for the expression of genes at high levels in the milk of transgenic animals. In particular, the present invention provides recombinant signal peptide sequences derived from a-lactalbumin and aS1-casein milk genes suitable for leading protein secretion in the mammary gland. These gene cassettes are capable of delivering different transgenic constructs which result in the production of full-length or B domain-deleted therapeutic levels of biologically active human FVIII in the transgenic animals in vivo. Within the scope of the invention are also method for producing the transgenic non-human mammal, such as mouse, rat, rabbit, goat, sheep, pig and bovine species, capable of expressing human FVIII, and methods of making milk and methods of identifying protein from the transgenic milk.

    摘要翻译： 血友病A是凝血因子VIII（FVIII）缺乏或异常引起的主要遗传性出血性疾病之一。 已经用全血浆或纯化的FVIII浓缩物治疗血友病患者。 传播血源性病毒的风险和高度纯化的FVIII的成本是限制血友病治疗预防的主要因素。 生物技术革命带来的挑战之一是开发大规模经济生产高纯度蛋白质的方法。 本发明提供了可用于在转基因动物的牛奶中高水平表达基因的改进的乳腺表达盒。 特别地，本发明提供了衍生自α-乳白蛋白的重组信号肽序列和适合于在乳腺中引导蛋白质分泌的α1酪蛋白乳基因。 这些基因盒能够递送不同的转基因构建体，其导致在转基因动物体内产生全长或B结构域缺失的生物活性人类FVIII的治疗水平。 在本发明的范围内，也是用于生产能够表达人FVIII的转基因非人哺乳动物如小鼠，大鼠，兔，山羊，绵羊，猪和牛物种的方法，以及制备牛奶的方法和鉴定方法 来自转基因乳的蛋白质。