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    • 118. 发明公开
    • Process for the extraction and purification of human recombinant gamma interferon
    • Verfahren zur Extraktion und Reinigung von menschlichem rekombinantem gamma-Interferon。
    • EP0468393A1
    • 1992-01-29
    • EP91112215.8
    • 1991-07-22
    • SCLAVO S.p.A.
    • Protasi, OtelloMannucci, Donatella
    • C07K1/36C07K14/555C12P21/02A61K38/21
    • C07K14/57A61K38/00
    • An extraction and purification process for human gamma interferon (HuIFN-gamma) from recombinant host cells producing the same in insoluble form is described, which comprises suspending the cells in a buffer solution having pH from 6.0 to 8.5, optionally complemented with an osmoprotector; disrupting the cells in conditions which do not activate the endocellular proteases; separating and solubilizing the pellet containing the insoluble HuIFN-gamma by extraction with phosphate buffer having pH comprised between 6.5 and 8.5 and finally purifying the solubilized HuIFN-gamma by one or more chromatographic techniques. Such a process allows non-degraded HuIFN-gamma to be obtained, with a biological activity and chemical-physical properties unaffected vis-a-vis the native product and in high yield and purity.
    • 描述了从产生不溶形式的重组宿主细胞的人γ-干扰素(HuIFN-γ)的提取和纯化方法,其包括将细胞悬浮在pH 6.0至8.5的缓冲溶液中,任选地与渗透保护剂互补; 在不激活内细胞蛋白酶的条件下破坏细胞; 通过用pH为6.5-8.5的磷酸盐缓冲液萃取,通过用一种或多种色谱技术纯化溶解的HuIFN-γ,分离并溶解含有不溶性HuIFN-γ的沉淀物。 这种方法允许获得未降解的HuIFN-γ,其生物活性和化学物理性质不受天然产物的影响,并且以高产率和纯度。