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    • 16. 发明申请
    • Methods For Enhancing A Secretion Efficiency Of Recombinant Foreign Protein In Yeast Expression System
    • 用于增强酵母表达系统中重组外源蛋白的分泌效率的方法
    • US20100184136A1
    • 2010-07-22
    • US12305768
    • 2006-06-20
    • Hyung-Kwon LimSung-Geun Kim
    • Hyung-Kwon LimSung-Geun Kim
    • C12P21/00
    • C12N15/81
    • Provided is a method for improving secretion efficiency of a recombinant foreign protein in a yeast expression system. The method comprises transforming a yeast host with a recombinant foreign gene construct comprising a galactose-inducible promoter, a secretion signal sequence and a gene encoding the foreign protein to construct a transformed yeast strain; and culturing the transformed yeast strain under the condition that the activity of the galactose-inducible promoter is controlled. Improved secretion efficiency of the foreign protein can be achieved by decreasing over-expression-induced insoluble precipitation of the recombinant foreign protein suffered by a conventional galactose-inducible promoter-based yeast expression system, via appropriate control of a level of galactose functioning as an inducer of the galactose-inducible promoter in cells. Due to improved secretion efficiency of the recombinant foreign protein, present invention makes a contribution to improvement in productivity of recombinant foreign proteins in the yeast expression system and reduction in production costs.
    • 提供了一种用于提高酵母表达系统中重组外源蛋白的分泌效率的方法。 该方法包括用包含半乳糖诱导型启动子,分泌信号序列和编码外源蛋白的基因的重组外源基因构建体来转化酵母宿主以构建转化的酵母菌株; 并在控制半乳糖诱导型启动子的活性的条件下培养转化的酵母菌株。 通过适当控制用作诱导剂的半乳糖水平,通过减少由常规的基于半乳糖诱导型启动子的酵母表达系统所承受的重组外源蛋白质的过表达诱导的不溶性沉淀,可以实现外源蛋白质的分泌效率的提高。 的半乳糖诱导型启动子。 由于重组外源蛋白的分泌效率提高,本发明对于提高酵母表达系统中重组外源蛋白质的生产力和生产成本的降低有所贡献。
    • 18. 发明申请
    • Mutant of Granulocyte-Colony Stimulating Factor (G-Csf) and Chemically Conjugated Polypeptide Thereof
    • 粒细胞集落刺激因子(G-Csf)和化学结合多肽的突变体
    • US20080200657A1
    • 2008-08-21
    • US11995968
    • 2006-07-19
    • Kwan-Yub KangJeong-Woon Hong
    • Kwan-Yub KangJeong-Woon Hong
    • C07K14/435
    • C07K14/535A61K47/60
    • Provided are mutants of human granulocyte-colony stimulating factor (G-CSF) designed for specific chemical conjugation, and chemical conjugates thereof for use as an adjuvant in the treatment of cancer. The present invention provides a mutant of a G-CSF in which a threonine (Thr) residue at position 133 of G-CSF comprising the amino acid sequence identified in SEQ ID NO: 1 is substituted with a cysteine (Cys) residue. In addition, the invention provides a mutant of a G-CSF in which a cysteine (Cys) residue is inserted between a glycine (Gly) residue at position 135 and an alanine (Ala) residue at position 136 of G-CSF. Further, the invention provides a chemically conjugated mutant G-CSF to which biocompatible polymer such as polyethylene glycol (PEG) was attached at the cysteine residue, which was introduced by the substitution or insertion mutation, increasing its in vivo retention time without reducing in vivo biological activity due to the conjugation with the biocompatible polymer, thereby ultimately extending the in vivo biological activity.
    • 提供设计用于特定化学共轭的人粒细胞集落刺激因子(G-CSF)的突变体,以及用作癌症治疗中的佐剂的化学共轭物。 本发明提供了G-CSF的突变体,其中包含SEQ ID NO:1中鉴定的氨基酸序列的G-CSF的第133位的苏氨酸(Thr)残基被半胱氨酸(Cys)残基取代。 此外,本发明提供了G-CSF的突变体,其中将半胱氨酸(Cys)残基插入在135位的甘氨酸(Gly)残基和G-CSF的第136位的丙氨酸(Ala)残基之间。 此外,本发明提供了在半胱氨酸残基处连接生物相容性聚合物如聚乙二醇(PEG)的化学偶联的突变体G-CSF,其通过取代或插入突变引入,增加其体内保留时间而不降低体内 由于与生物相​​容性聚合物共轭而产生的生物活性,从而最终延长了体内的生物活性。