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    • 3. 发明授权
    • Analysis method using reporter (label) intermolecular interaction
    • 分析方法采用报告(标记)分子间相互作用
    • US07595202B2
    • 2009-09-29
    • US10475540
    • 2002-04-23
    • Teruyuki NagamuneHiroshi UedaYoshiyuki OhiroNorio Shibata
    • Teruyuki NagamuneHiroshi UedaYoshiyuki OhiroNorio Shibata
    • G01N33/541G01N21/76G01N33/533G01N33/566G01N33/53G01N33/542
    • G01N33/54306G01N33/533G01N33/542G01N33/582
    • The present invention provides a method for controlling the spatial arrangement of an energy donor and an energy acceptor in a reaction complex in order to carry out the measurement with good precision and high sensitivity, and a measurement system using the method. Two types of material having affinity for the subject material to be measured are respectively labeled with a combination of reporters that give rise to energy transfer; those that have been thus obtained by labeling these materials are each further labeled with materials that have weak affinity for each other to give reagents, which are then mixed with a sample to give the reaction complex. Each of the materials is brought into spatial proximity by binding based on the affinity among the materials having weak affinity for each other in the reaction complex, and since that condition is stably maintained, a more efficient energy transfer occurs.
    • 本发明提供了一种用于控制反应复合物中能量供体和能量受体的空间排列的方法,以便以高精度和高灵敏度进行测量,以及使用该方法的测量系统。 分别用引起能量转移的记者的组合来标示对被测量物质具有亲和性的两种类型的材料; 通过标记这些材料得到的那些物质每个进一步用彼此具有弱亲和力的材料进行标记,得到试剂,然后将其与样品混合以得到反应络合物。 通过基于在反应络合物中彼此之间具有弱亲和力的材料之间的亲和力,通过结合使每种材料变得空间接近,并且由于稳定地保持该条件,因此发生更有效的能量转移。
    • 4. 发明申请
    • Method of assaying interaction between proteins
    • 测定蛋白质之间相互作用的方法
    • US20060252028A1
    • 2006-11-09
    • US10524564
    • 2003-08-15
    • Hiroshi UedaTeruyuki Nagamune
    • Hiroshi UedaTeruyuki Nagamune
    • C12Q1/70C07H21/04C12P21/06C12N1/21C07K16/18C12N15/74
    • C40B40/02C12N15/1037C12N15/1055C12N15/70
    • A novel method for determining interaction between VH fragment and VL fragment of the variable region of an antibody is provided by the present invention. The method according to this invention can be widely used for detection of protein interactions. If a phagemid vector containing an amber codon is used for transformation of an amber suppressor strain of E. coli according to the method of the invention to produce phages, both of the VH fragment and the VL fragment will be displayed on the phage particles. In contrast, if the same vector is used to transformation of an amber non-suppressing strain of E. coli to produce phages, for the presence of the amber codon, only the VH fragment will be displayed on the phage particles, while the VL fragment will be secreted into the culture medium by the E. coli. Thus, display switch occurs. The interaction between the VH fragment and the VL fragment can be determined, by immobilizing the VL fragment secreted into the culture medium on a solid phase and quantifying binding between the VH fragment displayed by phages and the VL fragment.
    • 本发明提供了用于确定抗体可变区的VH片段和VL片段之间相互作用的新方法。 根据本发明的方法可广泛用于蛋白质相互作用的检测。 如果含有琥珀密码子的噬菌体载体用于根据本发明的方法转化大肠杆菌的琥珀抑制菌株以产生噬菌体,那么VH片段和VL片段都将显示在噬菌体颗粒上。 相比之下,如果使用相同的载体转化大肠杆菌的琥珀非抑制菌株以产生噬菌体,则对于琥珀密码子的存在,只有VH片段将显示在噬菌体颗粒上,而VL片段 将被大肠杆菌分泌到培养基中。 因此,显示切换发生。 可以通过将分泌到培养基中的VL片段固定在固相上并定量噬菌体展示的VH片段与VL片段之间的结合来确定VH片段和VL片段之间的相互作用。
    • 8. 发明申请
    • FLUOROIMMUNOASSAY METHOD
    • 荧光分光法
    • US20120270338A1
    • 2012-10-25
    • US13510105
    • 2010-11-19
    • Hiroshi UedaRyoji AbeMasaki IharaHiroaki Takagi
    • Hiroshi UedaRyoji AbeMasaki IharaHiroaki Takagi
    • G01N21/76C07K17/02
    • G01N33/582A61K49/0058C07K16/18C07K16/44C07K2317/56C07K2317/622C07K2317/92G01N21/6428G01N33/6857G01N2021/6439
    • An object is to provide an immunoassay method requiring neither a solid-phase immobilization step nor a washing step, enabling quick and simple quantitative measurement of a target substance in a liquid phase and capable of visualizing an antigen. Such an object is attained by measuring the concentration of a target antigen present in a test substance by sequentially performing a step (a) of bringing an antibody light-chain variable region polypeptide and an antibody heavy-chain variable region polypeptide labeled with a fluorescent dye into contact with an antigen in a test substance in a liquid phase; or bringing an antibody heavy-chain variable region polypeptide and an antibody light-chain variable region polypeptide labeled with a fluorescent dye into contact with an antigen in a test substance in a liquid phase; a step (b) of measuring the fluorescence intensity of the fluorescent dye; and a step (c) of computationally obtaining the level of the antigen contained in the test substance with reference to a positive correlation between the concentration of the antigen in a liquid phase and the fluorescence intensity of the fluorescent dye.
    • 本发明的目的是提供一种不需要固相固定步骤和洗涤步骤的免疫测定方法,能够快速和简单地定量测定液相中的目标物质并能够使抗原呈现。 通过依次进行使荧光染料标记的抗体轻链可变区多肽和抗体重链可变区多肽的步骤(a),测定待测物质中存在的靶抗原的浓度, 与液相中测试物质中的抗原接触; 或使用荧光染料标记的抗体重链可变区多肽和抗体轻链可变区多肽与液相中的测试物质中的抗原接触; 测量荧光染料的荧光强度的步骤(b); 和(c)参考在液相中的抗原浓度与荧光染料的荧光强度之间的正相关性来计算获得测试物质中含有的抗原水平的步骤(c)。