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    • 7. 发明授权
    • Method of purification of human BCDF
    • 人BCDF纯化方法
    • US5610284A
    • 1997-03-11
    • US275663
    • 1994-07-15
    • Daisuke EjimaYutaka SatoMayumi WatanabeMasayo DateYoshiyuki Takahara
    • Daisuke EjimaYutaka SatoMayumi WatanabeMasayo DateYoshiyuki Takahara
    • A61K38/00C07K14/54C07K1/14C12N15/19
    • C07K14/5412A61K38/00
    • There are disclosed (i) a purification process for obtaining a human BCDF having the intramolecular disulfide linkage and the stereostructure of natural type human BCDF which comprises subjecting to an oxidation reaction and a refolding treatment a reduced type human BCDF obtained by culturing a microorganism having a human BCDF gene integrated therein and solubilized with guanidine hydrochloride, characterized in that after the oxidation reaction, a gel filtration chromatographic treatment is conducted under the conditions of the guanidine hydrochloride concentration adjusted to 4-7M; (ii) a purification process for obtaining a natural type human BCDF monomer by removing the organic solvent from an organic solvent-containing solution of human BCDF, characterized in that the solution is passed through a gel filtration chromatographic column equilibrated with an organic solvent, followed by eluting according to a stepwise or linear gradient program; and (iii) a human BCDF purification process comprising an ion exchange chromatographic treatment and a reversed phase HPLC step, in combination (i) or (ii). According to these purification processes, it becomes possible to remove the impurities derived from a microorganism and human BCDF analogs, and the thus obtained natural type human BCDF has a high purity and can be utilized for pharmaceutical preparations.
    • 公开了(i)用于获得具有分子内二硫键的人BCDF和天然型人BCDF的立体结构的纯化方法,其包括进行氧化反应和重折叠处理,所述还原型人BCDF通过培养具有 人BCDF基因整合在其中并用盐酸胍溶解,其特征在于,在氧化反应后,在盐酸胍浓度调节至4-7M的条件下进行凝胶过滤色谱处理; (ii)通过从含有机溶剂的人BCDF中除去有机溶剂来获得天然型人BCDF单体的纯化方法,其特征在于使溶液通过用有机溶剂平衡的凝胶过滤色谱柱,随后 通过逐步或线性梯度程序洗脱; 和(iii)包含离子交换色谱处理和反相HPLC步骤(组合(i)或(ii))的人BCDF纯化方法。 根据这些纯化方法,可以除去由微生物和人BCDF类似物衍生的杂质,由此得到的天然型人BCDF具有高纯度,可用于药物制剂。