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14 条记录 (耗时 0.016 秒)

    USE OF VALPROIC ACID FOR ENHANCING PRODUCTION OF RECOMBINANT PROTEINS IN MAMMALIAN CELLS
    1.
    发明申请
    USE OF VALPROIC ACID FOR ENHANCING PRODUCTION OF RECOMBINANT PROTEINS IN MAMMALIAN CELLS 审中-公开
    用于增强在马马里亚细胞中重组蛋白生产的有效性

    公开(公告)号：US20090023186A1

    公开(公告)日：2009-01-22

    申请号：US11781252

    申请日：2007-07-22

    申请人： Markus Hildinger ,  Gaurav Backliwal ,  Florian Wurm

    发明人： Markus Hildinger ,  Gaurav Backliwal ,  Florian Wurm

    IPC分类号： C12P21/00

    CPC分类号： C12P21/02 ,  C07K16/00

    摘要： Culturing cells for the commercial production of proteins for diagnosis and therapy is a costly and time consuming process. The equipment required is expensive, and production cost are high. In order to provide commercially viable processes it is desirable to use cell lines which produce large quantities of product with each production run. However, most cells do not produce large quantities of desired product per se either because they do not produce a large quantity of product per unit of time (specific productivity) or because they do not survive long enough in the culture medium (time). Here, we identified that addition of a valproic acid compound to the culture medium increases overall (batch) yield and titer. More importantly, compared to the widely used sodium butyrate, batch yields using a valproic acid compound as a medium additive are significantly higher.

    摘要翻译： 用于商业生产蛋白质用于诊断和治疗的培养细胞是昂贵且耗时的过程。 所需设备价格昂贵，生产成本高。 为了提供商业可行的方法，期望使用在每次生产运行时产生大量产品的细胞系。 然而，大多数细胞本身并不产生大量的所需产品，因为它们每单位时间（比生产率）不产生大量的产品，或者因为它们在培养基（时间）内不能足够长的时间生存。 在这里，我们确定向培养基中加入丙戊酸化合物提高了总体（批）产率和滴度。 更重要的是，与广泛使用的丁酸钠相比，使用丙戊酸化合物作为中等添加剂的批产量显着更高。

    DECREASING GENE EXPRESSION IN A MAMMALIAN SUBJECT IN VIVO VIA AAV-MEDIATED RNAi EXPRESSION CASSETTE TRANSFER
    3.
    发明申请
    DECREASING GENE EXPRESSION IN A MAMMALIAN SUBJECT IN VIVO VIA AAV-MEDIATED RNAi EXPRESSION CASSETTE TRANSFER 审中-公开
    通过AAV介导的RNAi表达载体转移，在VIVO中降低MAMMALIAN受体中的基因表达

    公开(公告)号：US20050019927A1

    公开(公告)日：2005-01-27

    申请号：US10604340

    申请日：2003-07-13

    申请人： Markus Hildinger ,  Alberto Auricchio

    发明人： Markus Hildinger ,  Alberto Auricchio

    IPC分类号： A61K38/00 ,  A61K48/00 ,  C12N5/02 ,  C12N15/113 ,  C12N15/864 ,  C12N15/86

    CPC分类号： C12N15/113 ,  A61K38/00 ,  A61K48/00 ,  C12N15/86 ,  C12N2310/111 ,  C12N2310/14 ,  C12N2750/14143 ,  C12N2840/20

    摘要： Decreasing the expression of genes in a mammalian subject has multiple applications ranging from cancer therapy to anti-infective therapy or treatment of autosomal dominant genetic disorders. Yet, there is still a lack of efficient technologies to achieve that goal in mammalian subjects in vivo. The present invention relates to methods for decreasing gene expression by administering to a mammalian subject a recombinant adeno-associated viral vector in vivo with said vector comprising an RNA interference (RNAi) expression cassette whose RNA expression products directly or indirectly lead to a decrease in expression of the corresponding RNAi target gene. Upon successful transduction with the recombinant adeno-associated viral vector, the RNA expression products of the RNAi expression cassette will decrease the cellular concentration of the mRNA transcripts of the RNAi target gene, thus resulting in decreased concentration of the protein encoded by the RNAi target gene.

    摘要翻译： 减少哺乳动物受试者中基因的表达具有多种应用，从癌症治疗到抗感染治疗或治疗常染色体显性遗传疾病。 然而，在哺乳动物体内仍然缺乏有效的技术来实现这一目标。 本发明涉及降低基因表达的方法，其通过向哺乳动物受试者施用重组腺相关病毒载体，所述载体包含RNA干扰（RNAi）表达盒，其RNA表达产物直接或间接导致表达降低 的相应RNAi靶基因。 在用重组腺相关病毒载体成功转导后，RNAi表达盒的RNA表达产物将降低RNAi靶基因的mRNA转录物的细胞浓度，从而导致由RNAi靶基因编码的蛋白质的浓度降低 。

    Super-size adeno-associated viral vector harboring a recombinant genome larger than 5.7 kb
    4.
    发明授权
    Super-size adeno-associated viral vector harboring a recombinant genome larger than 5.7 kb 失效
    超大尺寸腺相关病毒载体携带大于5.7kb的重组基因

    公开(公告)号：US07943374B2

    公开(公告)日：2011-05-17

    申请号：US11161890

    申请日：2005-08-21

    申请人： Markus Hildinger

    发明人： Markus Hildinger

    IPC分类号： C12N15/63 ,  C12N15/85

    CPC分类号： C12N15/86 ,  A61K48/00 ,  C12N2750/14143

    摘要： Prior art teaches an effective packaging capacity for adeno-associated virus and adeno-associated viral vectors of 4.1 kb to 4.9 kb as well as a packaging limit of 5.2 kb to 5.6 kb. However, the inventor discovered that this packaging limit as well as that effective packaging capacity does not apply to all AAV serotypes: Whereas it is true that efficient packaging of AAV serotype 2 is limited to less than 5 kb, the inventor discovered that one can efficiently package more than 6 kb of genetic information into AAV capsids of other AAV serotypes, particularly into capsids of AAV serotype 5 and—to a lesser extent—into capsids of AAV serotype 7. This discovery will be useful in the context of gene therapy where large transgenes will have to be transferred such as the ABCA4 coding sequence, the Factor VIII coding sequence, the B-deleted Factor VIII coding sequence or minidystrophin genes.

    摘要翻译： 现有技术教导了4.1kb至4.9kb的腺相关病毒和腺相关病毒载体的有效包装能力以及5.2kb至5.6kb的包装限制。 然而，发明人发现，这种包装限制以及有效的包装能力并不适用于所有AAV血清型：鉴于AAV血清型2的有效包装确实被限制在小于5kb，发明人发现可以有效地 将超过6kb的遗传信息包装到其他AAV血清型的AAV衣壳中，特别是AAV血清型5的衣壳中，并且在较小程度上包装到AAV血清型7的衣壳中。该发现将在基因治疗的背景下有用，其中大的 必须转移转基因，如ABCA4编码序列，因子VIII编码序列，缺失B因子VIII编码序列或微小营养不良蛋白基因。

    CELL CULTIVATION AND PRODUCTION OF RECOMBINANT PROTEINS BY MEANS OF AN ORBITAL SHAKE BIOREACTOR SYSTEM WITH DISPOSABLE BAGS AT THE 1,500 LITER SCALE
    9.
    发明申请
    CELL CULTIVATION AND PRODUCTION OF RECOMBINANT PROTEINS BY MEANS OF AN ORBITAL SHAKE BIOREACTOR SYSTEM WITH DISPOSABLE BAGS AT THE 1,500 LITER SCALE 审中-公开
    细胞培养和生产重组蛋白质的方法是通过在1,500 LITER SCALE上的具有可拆卸袋的ORBITAL SHAKE生物反应器系统

    公开(公告)号：US20090233334A1

    公开(公告)日：2009-09-17

    申请号：US12046374

    申请日：2008-03-11

    申请人： Markus Hildinger ,  Florian Wurm ,  Matthieu Stettler ,  Maria DeJesus

    发明人： Markus Hildinger ,  Florian Wurm ,  Matthieu Stettler ,  Maria DeJesus

    IPC分类号： C12P21/04 ,  C12N7/00 ,  C12N5/04 ,  C12N5/06 ,  C12N5/02 ,  C12N5/08 ,  C12N1/00

    CPC分类号： C12P21/02 ,  C12M23/14 ,  C12M27/16 ,  C12M41/02

    摘要： The present invention provides a novel method for culturing cells as well as a novel method for producing a recombinant protein by culturing cells at large scale (up to 1,500 L nominal volume and 750 L working volume), whereby an inflated bag provides a sterile, disposable cultivation chamber. The inflated bag is partially filled with liquid cultivation media and cells, and placed into a containment vessel. The containment vessel is positioned onto an orbitally shaken platform. The orbital shaking moves the containment vessel and thus the bag and induces thereby motion to the liquid contained therein (“shake mixing”). This motion (caused by orbital shaking) induces a dynamic force field that ensures cell suspension, bulk mixing, and oxygen transfer from the liquid surface to the respiring cells without damaging shear or foam generation.

    摘要翻译： 本发明提供了一种培养细胞的新方法以及通过大规模培养细胞（达1,500L标称体积和750L工作体积）来生产重组蛋白的新方法，由此充气袋提供无菌的，一次性的 栽培室。 充气袋部分地装有液体培养基和细胞，并放置在容纳容器中。 安全壳位于轨道上摇摆的平台上。 轨道振荡使得容纳容器和袋子移动并引起其运动到其中容纳的液体（“混合”）。 这种运动（由轨道震荡引起）引起动态力场，确保细胞悬浮，本体混合和从液体表面到呼吸细胞的氧传递，而不损害剪切或泡沫产生。