Modified chitin binding domain and uses thereof
    2.
    发明授权
    Modified chitin binding domain and uses thereof 失效
    改性的几丁质结合结构域及其用途

    公开(公告)号:US06897285B2

    公开(公告)日:2005-05-24

    申请号:US10375913

    申请日:2003-02-26

    IPC分类号: C12N9/24 C07K14/00

    摘要: Compositions and methods are provided for reversibly binding chitin binding domain (CBD) to a chitin or equivalent substrate under non denaturing conditions. CBD is modified preferably by a mutation to achieve this change in properties. In one embodiment, an aromatic amino acid residue located in the binding cleft of the CBD was altered resulting in reversible binding affinity for substrate in select conditions. Creating a modified CBD with an altered binding affinity for substrate provides new uses for CBD not previously possible with unmodified CBD which binds irreversibly to chitin.

    摘要翻译: 提供了组合物和方法,用于在非变性条件下将壳多糖结合域(CBD)可逆地结合到壳多糖或等效底物。 CBD优选通过突变来改变以实现这种性质的变化。 在一个实施方案中,位于CBD的结合裂缝中的芳族氨基酸残基被改变,导致在选择的条件下对底物的可逆结合亲和力。 用改良的底物结合亲和力建立经修饰的CBD,为以前不可能与未结合的CBD结合的CBD提供了新的用途,后者不可逆地与几丁质结合。

    Methods and Compositions for Concentrating Secreted Recombinant Protein
    5.
    发明申请
    Methods and Compositions for Concentrating Secreted Recombinant Protein 失效
    浓缩分泌重组蛋白的方法和组合

    公开(公告)号:US20120107877A1

    公开(公告)日:2012-05-03

    申请号:US13346258

    申请日:2012-01-09

    IPC分类号: C12P21/00

    摘要: Methods and compositions are described that relate to obtaining concentrated preparations of secreted recombinant proteins. These proteins are expressed in the form of fusion proteins with a chitin-binding domain (CBD). The fusion proteins are capable of being concentrated in the presence of chitin. Also described is: a shuttle vector that includes a modified LAC4 promoter; a chitinase-negative host cell; a CBD capable of eluting from chitin under non-denaturing conditions; and sterilized chitin, which can be optionally magnetized for facilitating recovery of recombinant protein.

    摘要翻译: 描述了与获得分泌的重组蛋白质的浓缩制剂相关的方法和组合物。 这些蛋白质以融合蛋白的形式与几丁质结合结构域(CBD)表达。 融合蛋白能够在几丁质的存在下被浓缩。 还描述了:包含经修饰的LAC4启动子的穿梭载体; 几丁质酶阴性宿主细胞; 一种能够在非变性条件下从几丁质中洗脱的CBD; 和灭菌的几丁质,其可以任选地被磁化以促进重组蛋白的回收。

    Methods and Compositions for Concentrating Secreted Recombinant Proteins
    6.
    发明申请
    Methods and Compositions for Concentrating Secreted Recombinant Proteins 有权
    用于浓缩分泌重组蛋白的方法和组合物

    公开(公告)号:US20110071280A1

    公开(公告)日:2011-03-24

    申请号:US12399059

    申请日:2009-03-06

    IPC分类号: C07K1/107 B82Y5/00

    摘要: Methods and compositions are described that relate to obtaining concentrated preparations of secreted recombinant proteins. These proteins are expressed in the form of fusion proteins with a chitin-binding domain (CBD). The fusion proteins are capable of being concentrated in the presence of chitin. Also described is: a shuttle vector that includes a modified LAC4 promoter; a chitinase-negative host cell; a CBD capable of eluting from chitin under non-denaturing conditions; and sterilized chitin, which can be optionally magnetized for facilitating recovery of recombinant protein.

    摘要翻译: 描述了与获得分泌的重组蛋白质的浓缩制剂相关的方法和组合物。 这些蛋白质以融合蛋白的形式与几丁质结合结构域(CBD)表达。 融合蛋白能够在几丁质的存在下被浓缩。 还描述了:包含经修饰的LAC4启动子的穿梭载体; 几丁质酶阴性宿主细胞; 一种能够在非变性条件下从几丁质中洗脱的CBD; 和灭菌的几丁质,其可以任选地被磁化以促进重组蛋白的回收。

    Solubilization and Purification of a Target Protein Fused to a Mutant Maltose-Binding Protein
    8.
    发明申请
    Solubilization and Purification of a Target Protein Fused to a Mutant Maltose-Binding Protein 失效
    融合到突变体麦芽糖结合蛋白的靶蛋白的溶解和纯化

    公开(公告)号:US20120028305A1

    公开(公告)日:2012-02-02

    申请号:US13259837

    申请日:2009-04-01

    CPC分类号: C07K14/245 C07K2319/24

    摘要: Methods and compositions are provided that relate to a composition that includes a modified maltose-binding protein (MBP) which when fused to a protein results in an increase in binding affinity for maltodextrin compared with the wild type MBP fused to the protein, the modified MBP maintaining enhanced solubility, The modification includes a mutation selected from the group consisting of: F68L, I318V, Q326R, V344M, and T372TTTITITTTLGIEGR387 or consist of two or more mutations selected from the group consisting of: F68L, S146T, A313V, I318V, I318A, Q326R, V344M and T372TTTITITTTLGIEGR387 mutants.

    摘要翻译: 提供了与包含修饰的麦芽糖结合蛋白(MBP)的组合物相关的方法和组合物,当与蛋白质融合时,与蛋白质融合的野生型MBP相比,与蛋白质融合导致与麦芽糖糊精结合亲和力的增加,修饰的MBP 维持增加的溶解度。该修饰包括选自以下的突变:F68L,I318V,Q326R,V344M和T372TTTITITTTLGIEGR387,或由两个或更多个选自F68L,S146T,A313V,I318V,I318A, Q326R,V344M和T372TTTITITTTLGIEGR387突变体。

    Methods and compositions for concentrating secreted recombinant protein
    9.
    发明授权
    Methods and compositions for concentrating secreted recombinant protein 失效
    用于浓缩分泌的重组蛋白的方法和组合物

    公开(公告)号:US07517671B2

    公开(公告)日:2009-04-14

    申请号:US11242553

    申请日:2005-10-03

    IPC分类号: C12P21/02

    摘要: Methods and compositions are described that relate to obtaining concentrated preparations of secreted recombinant proteins. These proteins are expressed in the form of fusion proteins with a chitin-binding domain (CBD). The fusion proteins are capable of being concentrated in the presence of chitin. Also described is: a shuttle vector that includes a modified LAC4 promoter; a chitinase-negative host cell; a CBD capable of eluting from chitin under non-denaturing conditions; and sterilized chitin, which can be optionally magnetized for facilitating recovery of recombinant protein.

    摘要翻译: 描述了与获得分泌的重组蛋白质的浓缩制剂相关的方法和组合物。 这些蛋白质以融合蛋白的形式与几丁质结合结构域(CBD)表达。 融合蛋白能够在几丁质的存在下被浓缩。 还描述了:包含经修饰的LAC4启动子的穿梭载体; 几丁质酶阴性宿主细胞; 一种能够在非变性条件下从几丁质中洗脱的CBD; 和灭菌的几丁质,其可以任选地被磁化以促进重组蛋白的回收。