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    • 5. 发明公开
    • METHOD FOR DETECTING SUBSTANCE IN BIOLOGICAL SAMPLE
    • 爱尔兰生物科学研究所的VERFAHRENFÜRDEN NACHWEIS EINES STOFFES
    • EP2416158A1
    • 2012-02-08
    • EP10758802.2
    • 2010-03-31
    • Japan Tobacco, Inc.Virus Ikagaku Kenkyusho Inc.
    • TAKAKURA, YoshimitsuOKA, NaomiKONDO, Kazuhiro
    • G01N33/543C07K17/00
    • G01N33/54393G01N33/54306G01N33/54353G01N33/56994G01N33/6854G01N2333/035G01N2333/36
    • The present invention provides a method for detecting a substance in a biological sample, a carrier for using in the method, and a kit. The method of the present invention includes 1) providing a carrier on which a biotin-binding protein is bound and providing a biotinylated protein by biotinylating a protein that specifically binds to a substance to be detected; 2) binding the biotinylated protein to the carrier provided in step 1) to produce a biotinylated protein-bound carrier; 3) mixing (a) a biological sample, and (b-i) a cell homogenate extract prepared from cells of the same species as that of the host cells used for expressing, for example, the biotin-binding protein in step 1), and a biotin-binding protein, or (b-ii) a cell homogenate extract prepared from cells of the same species as that of the host cells used for expressing, for example, the biotin-binding protein in step 1) and genetically engineered to express a biotin-binding protein, and adding the mixture to the biotinylated protein-bound carrier produced in step 2); and 4) detecting the substance specifically bound to the biotinylated protein.
    • 本发明提供了用于检测生物样品中物质的方法,该方法中使用的载体和试剂盒。 本发明的方法包括:提供结合有生物素结合蛋白的载体,并通过生物素化与特异性结合待检测物质的蛋白质提供生物素化蛋白; 2)将生物素化蛋白质与步骤1)中提供的载体结合以产生生物素化的蛋白质结合的载体; 3)混合(a)生物样品和(bi)由与用于在步骤1)中表达例如生物素结合蛋白的宿主细胞相同种类的细胞制备的细胞匀浆提取物,和 生物素结合蛋白,或(b-ii)从与用于表达例如生物素结合蛋白的步骤1中所用的宿主细胞相同种类的细胞制备的细胞匀浆提取物),并进行遗传工程化以表达 生物素结合蛋白,并将混合物加入步骤2)中产生的生物素化蛋白质结合的载体上; 和4)检测与生物素化蛋白质特异结合的物质。
    • 6. 发明公开
    • METHOD FOR DETECTING SUBSTANCE IN BIOLOGICAL SAMPLE
    • 爱尔兰生物科学研究所的VERFAHRENFÜRDEN NACHWEIS EINES STOFFES
    • EP2405268A1
    • 2012-01-11
    • EP10748752.2
    • 2010-03-02
    • Japan Tobacco, Inc.Virus Ikagaku Kenkyusho Inc.
    • TAKAKURA, YoshimitsuOKA, NaomiKONDO, Kazuhiro
    • G01N33/543G01N33/53C12N15/09
    • G01N33/54393
    • The present invention provides a method for detecting a substance in a biological sample, a carrier for using in the method, and a kit. The method of the present invention according to an embodiment includes:
      1) providing a carrier on which biotin is bound and providing a fusion protein between a protein that specifically binds to a substance to be detected and a biotin-binding protein;
      2) binding the fusion protein to the carrier provided in step 1) through binding between biotin and the biotin-binding protein to produce a fusion protein-bound carrier;
      3) mixing
      (a) a biological sample, and
      (b-i) a cell homogenate extract prepared from cells of the same species as the host cells used for expressing the fusion protein in step 1), and a biotin-binding protein, or
      (b-ii) a cell homogenate extract prepared from cells of the same species as the host cells used for expressing the fusion protein in step 1), and genetically engineered to express a biotin-binding protein,

      and adding the mixture to the fusion protein-bound carrier produced in step 2); and
      4) detecting a substance that has bound to the protein that specifically binds to a substance to be detected in the fusion protein.
    • 本发明提供了用于检测生物样品中物质的方法,该方法中使用的载体和试剂盒。 根据一个实施方案的本发明的方法包括:1)提供其上结合有生物素的载体,并在特异性结合待检测物质的蛋白质与生物素结合蛋白质之间提供融合蛋白; 2)通过生物素与生物素结合蛋白之间的结合将融合蛋白与步骤1)中提供的载体结合以产生融合蛋白结合的载体; 3)混合(a)生物样品和(bi)从与步骤1)中用于表达融合蛋白的宿主细胞相同种类的细胞制备的细胞匀浆提取物和生物素结合蛋白,或(b -ii)从与步骤1)中用于表达融合蛋白的宿主细胞相同物种的细胞制备的细胞匀浆提取物,并进行遗传工程改造以表达生物素结合蛋白,并将该混合物加入融合蛋白结合 载体在步骤2)中生产; 以及4)检测与融合蛋白中特异性结合待检测物质的蛋白质结合的物质。