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    • 42. 发明授权
    • Combinatorial DNA library for producing modified N-glycans in lower eukaryotes
    • 用于在低等真核生物中生产修饰的N-聚糖的组合DNA文库
    • US08883483B2
    • 2014-11-11
    • US13156804
    • 2011-06-09
    • Tillman U. GerngrossStefan WildtByung-kwon ChoiJuergen NettPiotr BobrowiczStephen HamiltonRobert Davidson
    • Tillman U. GerngrossStefan WildtByung-kwon ChoiJuergen NettPiotr BobrowiczStephen HamiltonRobert Davidson
    • C12N15/04C12N1/16C07K14/47C12N9/10C12N15/10C12N15/79
    • C12P21/005C07K14/47C12N9/1051C12N15/1082C12N15/79
    • The present invention relates to eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The invention provides nucleic acid molecules and combinatorial libraries which can be used to successfully target and express mammalian enzymatic activities such as those involved in glycosylation to intracellular compartments in a eukaryotic host cell. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified oligosaccharides are created or selected. N-glycans made in the engineered host cells have a Man5GlcNAc2 core structure which may then be modified further by heterologous expression of one or more enzymes, e.g., glycosyltransferases, sugar transporters and mannosidases, to yield human-like glycoproteins. For the production of therapeutic proteins, this method may be adapted to engineer cell lines in which any desired glycosylation structure may be obtained.
    • 本发明涉及具有修饰的寡糖的真核宿主细胞,其可以通过异源表达一组糖基转移酶,糖转运体和甘露糖苷酶进一步修饰,以成为用于产生哺乳动物例如人治疗性糖蛋白的宿主菌株。 本发明提供核酸分子和组合文库,其可用于成功靶向和表达哺乳动物酶活性,例如参与糖基化的真核宿主细胞中的细胞内区室的那些。 该方法提供了可用于表达和靶向参与糖基化的任何所需基因的工程化宿主细胞。 建立或选择具有修饰寡糖的宿主细胞。 在工程化宿主细胞中制备的N-聚糖具有Man5GlcNAc2核心结构,然后可以通过异源表达一种或多种酶,例如糖基转移酶,糖转运蛋白和甘露糖苷酶来进一步修饰,以产生人样糖蛋白。 为了生产治疗性蛋白质,该方法可适用于工程化可能获得任何所需糖基化结构的细胞系。
    • 47. 发明申请
    • Method and Apparatus For Pulse Welding
    • 脉冲焊接方法与装置
    • US20070235434A1
    • 2007-10-11
    • US11278344
    • 2006-03-31
    • Robert DavidsonDavid AlmyRichard Schuh
    • Robert DavidsonDavid AlmyRichard Schuh
    • B23K9/09
    • B23K9/092B23K9/093
    • A method and system for pulse welding provides an output pulse waveform. The waveform has at least a frequency from a range of available frequencies not limited to harmonic frequencies. The waveform provides power suitable for welding, and has a plurality of background portions alternating with a plurality of peak portions. A transition down occurs from the peak portion to the background portion with a first acceleration and a transition up occurs from the background portion to the peak portion with a second acceleration. At least one of the first and second accelerations are non-zero over at least most of the transition. The waveform is preferably created by switching an inverter. Both accelerations may be non-zero, and each transition may have two accelerations, one negative closer to the peak, and one positive closer to the background. The accelerations may be constant with opposite polarity. Preferably the process is current controlled during the transitions. The process frequency may be fixed or varying.
    • 脉冲焊接的方法和系统提供输出脉冲波形。 该波形至少有一个频率范围内的可用频率不限于谐波频率。 波形提供适合于焊接的功率,并且具有与多个峰部交替的多个背景部分。 从第一加速度的峰部分到背景部分发生转变下降,并且以第二加速度从背景部分到峰部分发生转变。 第一和第二加速度中的至少一个在至少大部分转换中都是非零的。 该波形优选地通过切换逆变器来产生。 两个加速度可以是非零,并且每个转变可以具有两个加速度,一个靠近峰值的加速度,另一个加速度更靠近背景。 加速度可以是相反极性的恒定。 优选地,该过程在过渡期间是电流控制的。 过程频率可以是固定的或变化的。