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    • 3. 发明授权
    • Methods for separating recombinant proteins in aqueous two-phase systems
    • 在两相体系中分离重组蛋白的方法
    • US08329877B2
    • 2012-12-11
    • US12723356
    • 2010-03-12
    • Jesús M. Sanz MoralesBeatriz Maestro Garcia-DonasMiguel Arevalo RodriguezIsabel Velasco UmpierrezAngel Cebolla Ramirez
    • Jesús M. Sanz MoralesBeatriz Maestro Garcia-DonasMiguel Arevalo RodriguezIsabel Velasco UmpierrezAngel Cebolla Ramirez
    • A23J1/00C07K1/00C07K14/00C07K16/00C07K17/00
    • C07K1/145
    • The object of the present invention is a procedure for the distribution, separation and purification in aqueous solution of recombinant proteins, based on the utilization of polypeptides with choline affinity. The invention is based on a phenomenon consisting of that two aqueous solutions with determinated components can be mixed, being distributed finally in two phases with different density. The fusionated proteins to said polypeptides with choline affinity are preferably located in one of the phases, while most of the cell extract proteins tend to go to the opposite phase. After a series of washings for removing the rest of the not desired material, this location can be inverted through the addition of a soluble molecule with affinity by the polypeptide fusionated to the protein of interest. This procedure allows modulating at convenience the presence of the protein or polypeptide of interest in one phase or another, possibiliting its purification with a high yield and purity grade. The invention represents an economic and scalable way of recombinant protein separation labelled preferably with choline-binding domains.
    • 本发明的目的是基于利用胆碱亲合力的多肽来分配,分离和纯化重组蛋白质的水溶液的方法。 本发明是基于这样的现象,其中可以混合具有确定组分的两种水溶液,最终以不同的密度两相分配。 具有胆碱亲和力的所述多肽的融合蛋白优选位于一个相中,而大多数细胞提取物蛋白倾向于进入相反阶段。 在一系列用于除去其余不需要的材料的洗涤液之后,通过与融合到感兴趣的蛋白质的多肽的亲和力加入可溶性分子,可以反转该位置。 该方法允许在一个阶段或另一个阶段方便地调节感兴趣的蛋白质或多肽的存在,可能以高产率和纯度等级进行纯化。 本发明代表优选用胆碱结合结构域标记的重组蛋白分离的经济和可扩展的方法。
    • 4. 发明申请
    • METHODS FOR SEPARATING RECOMBINANT PROTEINS IN AQUEOUS TWO-PHASE SYSTEMS
    • 在两相系统中分离重组蛋白的方法
    • US20100234567A1
    • 2010-09-16
    • US12723356
    • 2010-03-12
    • Jesús M. SANZ MORALESBeatriz Maestro Garcia-DonasMiguel Arevalo RodriguezIsabel Velasco UmpierrezAngel Cebolla Ramirez
    • Jesús M. SANZ MORALESBeatriz Maestro Garcia-DonasMiguel Arevalo RodriguezIsabel Velasco UmpierrezAngel Cebolla Ramirez
    • C07K1/14
    • C07K1/145
    • The object of the present invention is a procedure for the distribution, separation and purification in aqueous solution of recombinant proteins, based on the utilization of polypeptides with choline affinity. The invention is based on a phenomenon consisting of that two aqueous solutions with determinated components can be mixed, being distributed finally in two phases with different density. The fusionated proteins to said polypeptides with choline affinity are preferably located in one of the phases, while most of the cell extract proteins tend to go to the opposite phase. After a series of washings for removing the rest of the not desired material, this location can be inverted through the addition of a soluble molecule with affinity by the polypeptide fusionated to the protein of interest. This procedure allows modulating at convenience the presence of the protein or polypeptide of interest in one phase or another, possibiliting its purification with a high yield and purity grade. The invention represents an economic and scalable way of recombinant protein separation labelled preferably with choline-binding domains.
    • 本发明的目的是基于利用胆碱亲合力的多肽来分配,分离和纯化重组蛋白质的水溶液的方法。 本发明是基于这样的现象,其中可以混合具有确定组分的两种水溶液,最终以不同的密度两相分配。 具有胆碱亲和力的所述多肽的融合蛋白优选位于一个相中,而大多数细胞提取物蛋白倾向于进入相反阶段。 在一系列用于除去其余不需要的材料的洗涤液之后,通过与融合到感兴趣的蛋白质的多肽的亲和力加入可溶性分子,可以反转该位置。 该方法允许在一个阶段或另一个阶段方便地调节感兴趣的蛋白质或多肽的存在,可能以高产率和纯度等级进行纯化。 本发明代表优选用胆碱结合结构域标记的重组蛋白分离的经济和可扩展的方法。