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    • 9. 发明申请
    • DETECTION PHENOTYPING AND QUANTITATION OF CELLS WITH MULTIMERIC BINDING REAGENT
    • 用多种结合试剂检测细胞的定位和定量
    • WO2017015064A1
    • 2017-01-26
    • PCT/US2016/042341
    • 2016-07-14
    • THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY
    • DAVIS, Mark M.HUANG, Jun
    • A61K38/00C12N15/09
    • G01N33/56972C07K14/00C07K14/36G01N2333/70539
    • Methods and compositions are provided for labeling cells according to the specificity of a receptor of interes by contacting with a multimeric binding reagent. The multimeric binding reagent is based on a "tetrameric scaffold protein", which tetrameric scaffold protein has little or no affinity for biotin, and which comprises a C-terminal cysteine residue on one or more, usually all four of the polypeptides in the tetramer. The scaffold protein is modified by biotinylation at the terminal cysteines, to provide the central portion of the multimeric binding reagent. The biotinylated tetramer is combined with a high affinity biotin-binding protein tetramer. The complex thus generated has unfilled biotin-binding sites that can accommodate up to twelve biotin-tagged specific binding reagents, and is useful in various affinity selection and detection formats.
    • 提供方法和组合物,以通过与多聚体结合试剂接触,根据内在受体的特异性标记细胞。 多聚体结合试剂基于“四聚体支架蛋白”,其四聚体支架蛋白对生物素几乎没有或没有亲和力,并且其在四聚体中的一个或多个,通常全部四个多肽中包含C末端半胱氨酸残基。 通过在末端半胱氨酸的生物素化修饰支架蛋白,以提供多聚体结合试剂的中心部分。 生物素化四聚体与高亲和力生物素结合蛋白四聚体组合。 由此产生的复合物具有可以容纳多达12个生物素标记的特异性结合试剂的未填充的生物素结合位点,并且可用于各种亲和力选择和检测形式。