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    Site-specific labeling of affinity tags in fusion proteins
    3.
    发明申请
    Site-specific labeling of affinity tags in fusion proteins 审中-公开
    融合蛋白中亲和标签的位点特异性标记

    公开(公告)号：US20140038856A1

    公开(公告)日：2014-02-06

    申请号：US13890137

    申请日：2013-05-08

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Kyle GEE ,  Aleksey RUKAVISHNIKOV ,  Courtenay HART ,  Richard HAUGLAND ,  Wai-Yee LEUNG ,  Wayne PATTON ,  Zhenjun DIWU

    IPC: G01N33/58

    CPC classification number: G01N33/52 ,  A61K49/0021 ,  A61K49/0039 ,  A61K49/0041 ,  A61K49/0052 ,  C07D311/12 ,  C07D311/16 ,  C07D311/18 ,  C07F5/022 ,  G01N33/533 ,  G01N33/582

    Abstract: The present invention provides methods and fluorescent compounds that facilitate detecting and labeling of a fusion protein by being capable of selectively binding to an affinity tag. The fluorescent compounds have the general formula A(B)n, wherein A is a fluorophore, B is a binding domain that is a charged chemical moiety, a protein or fragment thereof and n is an integer from 1-6 with the proviso that the protein or fragment thereof not be an antibody or generated from an antibody. The present invention provides specific fluorescent compounds and methods used to detect and label fusion proteins that contain a poly-histidine affinity tag. These compounds have the general formula A(L)m(B)n wherein A is a fluorophore, L is a linker, B is an acetic acid binding domain, m is an integer from 1 to 4 and n is an integer from 1 to 6. The acetic acid groups interact directly with the positively charged histidine residues of the affinity tag to effectively label and detect a fusion protein containing such an affinity tag when present in an acidic or neutral environment.

    Abstract translation: 本发明提供了通过能够选择性地结合亲和标签来促进融合蛋白的检测和标记的方法和荧光化合物。 荧光化合物具有通式A（B）n，其中A是荧光团，B是作为带电化学部分的结合结构域，其蛋白质或片段，n是1-6的整数，条件是 蛋白质或其片段不是抗体或从抗体产生。 本发明提供用于检测和标记含有多组氨酸亲和标签的融合蛋白的特异性荧光化合物和方法。 这些化合物具有通式A（L）m（B）n，其中A是荧光团，L是连接体，B是乙酸结合域，m是1至4的整数，n是1至 乙酸基团与亲和标签的带正电荷的组氨酸残基直接相互作用，以有效地标记和检测当存在于酸性或中性环境中时含有这种亲和标签的融合蛋白。

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