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    • 3. 发明专利
    • Prevention of disulfide bond reduction during recombinant production of polypeptide
    • 防止重组生产多肽期间减少结缔组织
    • JP2014129358A
    • 2014-07-10
    • JP2014016227
    • 2014-01-30
    • Genentech Incジェネンテック, インコーポレイテッド
    • KAO YUNG-HSIANGMELODY TREXLER SCHMIDTMICHAEL W LAIRDWONG RITA LDANIEL P HEWITT
    • C07K1/113C12N15/09
    • C07K16/2887A61K39/39591C07K1/14C07K2317/24
    • PROBLEM TO BE SOLVED: To provide a method and means for protection of disulfide bonds to produce a correctly folded polypeptide which has a requested biological activity during a recombinant production of a polypeptide with a disulfide bond, particularly a multi-chain polypeptide comprising an intra-chain disulfide bond such as an antibody.SOLUTION: The method for preventing a reduction of disulfide bond in a polypeptide expressed in a recombinant host cell comprises supplying an inhibitor of a thioredoxin or thioredoxin-like protein to uncollected or collecting culture medium of the recombinant host cell. According to this method, the thioredoxin inhibitor directly inhibits the thioredoxin, or specifically inhibits a thioredoxin reductase. The thioredoxin inhibitor is alkyl 2-imidazolyl disulfide or a naphthoquinone spiroketal derivative, or they are yellow beeswax glucose (ATG) or a gold thiomalic acid salt (ATM).
    • 要解决的问题:提供用于保护二硫键以产生正确折叠的多肽的方法和手段,所述多肽在重组生产具有二硫键的多肽期间具有所要求的生物学活性,特别是多链多肽, 链二硫键如抗体。解决方案:用于预防重组宿主细胞中表达的多肽中二硫键还原的方法包括将硫氧还蛋白或硫氧还蛋白样蛋白质的抑制剂供应给重组宿主的未收集或收集培养基 细胞。 根据该方法,硫氧还蛋白抑制剂直接抑制硫氧还蛋白,或者特异性抑制硫氧还蛋白还原酶。 硫氧还蛋白抑制剂是2-咪唑基二硫化烷基或萘醌螺螺旋酮衍生物,或它们是黄蜂蜡葡萄糖(ATG)或金硫代酸盐(ATM)。