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    • 5. 发明申请
    • PREPARATION OF CELL EXTRACT AND ITS APPLICATION FOR CELL-FREE PROTEIN SYSTHESIS
    • 细胞提取物的制备及其对细胞免疫蛋白质的应用
    • US20100216184A1
    • 2010-08-26
    • US12279835
    • 2007-02-15
    • Dong-Myung KimTae-Wan KimChang-Gil ParkCha-Yong Choi
    • Dong-Myung KimTae-Wan KimChang-Gil ParkCha-Yong Choi
    • C12P21/06
    • C12P21/00C12M47/06
    • Disclosed is a process for simply preparing cell extracts for use as a catalyst of cell-free protein synthesis by centrifugation, which improves cost effectiveness and productivity of cell-free protein synthesis. Specifically, a conventional process for preparing cell extracts comprises the complicated steps, i.e. cell culture, cell lysis, high-speed centrifugation, pre-incubation, dialysis and the like. In comparison, the cell lysate just obtained by centrifugation is directly applied to protein synthesis, thereby providing higher producibility and more consistent productivity of protein than the conventional process. Further, the cell extracts are prepared by the simple process to reduce the protein production cost and time by about 60% and about 80%, respectively.
    • 公开了简单地制备细胞提取物以用作通过离心的无细胞蛋白质合成的催化剂的方法,这提高了无细胞蛋白质合成的成本效率和生产率。 具体地,用于制备细胞提取物的常规方法包括复杂的步骤,即细胞培养,细胞裂解,高速离心,预孵育,透析等。 相比之下,通过离心获得的细胞裂解物直接应用于蛋白质合成,从而提供比常规方法更高的可生产性和更一致的蛋白质生产力。 此外,通过简单的方法制备细胞提取物,以将蛋白质生产成本和时间分别降低约60%和约80%。
    • 7. 发明授权
    • Enhanced in vitro synthesis of biological macromolecules using a novel ATP regeneration system
    • 使用新型ATP再生系统增强生物大分子的体外合成
    • US06168931A
    • 2001-01-02
    • US09270814
    • 1999-03-17
    • James R. SwartzDong-Myung Kim
    • James R. SwartzDong-Myung Kim
    • C12P2106
    • C12N9/1033C07K14/5255C12P21/00
    • Compositions and methods are provided for the enhanced in vitro synthesis of biological molecules where ATP is required for synthesis. Of particular interest is the synthesis of polymers, e.g. nucleic acids, polypeptides, and complex carbohydrates. A homeostatic system is used for production of ATP, where the required high energy phosphate bonds are generated in situ, e.g. through coupling with an oxidation reaction. The homeostatic energy source will typically lack high energy phosphate bonds itself, and will therefore utilize free phosphate in the reaction mix during generation of ATP. Since inorganic phosphate can be an inhibitory by-product of synthesis, the period of time when synthesis is maintained in vitro can be extended. The homeostatic energy source is provided in combination with an enzyme that catalyzes the creation of high energy phosphate bonds and with an enzyme that can use that high energy phosphate bond to regenerate ATP.
    • 提供了用于合成需要ATP的生物分子的增强的体外合成的组合物和方法。 特别令人感兴趣的是聚合物的合成,例如 核酸,多肽和复合碳水化合物。 使用稳态系统来生产ATP,其中所需的高能磷酸键是原位生成的。 通过与氧化反应的偶联。 稳态能源本身通常缺乏高能量磷酸键,因此在产生ATP期间将在反应混合物中利用游离磷酸盐。 由于无机磷酸盐可以是合成的抑制副产物,因此可以延长合成时间在体外保持的时间。 稳态能量源与催化高能磷酸键的酶的结合,以及能够使用高能磷酸键重新生成ATP的酶组合提供。