公开(公告)号：EP2700950B1

公开(公告)日：2020-07-22

申请号：EP12774636.0

申请日：2012-04-20

Applicant: Korea Advanced Institute of Science and Technology

Inventor： YOON, Tae-Young

IPC: G01N33/68 ,  G01N21/64 ,  G01N33/574 ,  G01Q30/14 ,  G01Q60/20

公开(公告)号：EP2700950A2

公开(公告)日：2014-02-26

申请号：EP12774636.0

申请日：2012-04-20

Applicant: Korea Advanced Institute of Science and Technology

Inventor： YOON, Tae-Young

IPC: G01N33/68 ,  G01N33/533 ,  G01N33/574 ,  G01N21/64

Abstract: A method of analyzing protein-protein interactions at a single molecular level is disclosed. The method of analyzing the interactions between first proteins and second proteins at the single molecular level includes: preparing at least two substrates, in which first protein-binding molecules that are biomolecules to be bound to the first proteins are attached to each of the substrates; inducing binding between the first proteins and the first protein-binding molecules on the first substrate and the second substrate, respectively, by supplying the first proteins included in the control group-cell to the first substrate among the two substrates and supplying the first proteins included in the experimental group-cell to the second substrate among the two substrates; supplying cell lysates of cells including the marker-tagged second proteins to the first substrate and the second substrate, respectively, when the first proteins and the first protein-binding molecules are bound to the first substrate and the second substrate, respectively; and comparing and analyzing the interactions between the first proteins and the second proteins on the first substrate and the second substrate in the supply of the cell lysates to the first substrate and the second substrate, respectively. For observing the interactions between the first proteins and the second proteins, the state of each cell and activation levels of the first proteins can be compared and analyzed by comparing after varying of a type of cells supplying the first proteins.

Abstract translation: 在单分子水平上分析蛋白质 - 蛋白质相互作用的方法是游离缺失盘。 在单分子水平上分析第一蛋白质和第二蛋白质之间的相互作用的方法，包括：在哪个第一蛋白质结合分子做准备至少两个衬底是生物分子被束缚于所述第一蛋白质附着于各基板的; 诱导所述第一蛋白质和第一基板上的第一蛋白结合分子和所述第二基板之间，分别和结合时，可提供包括在控制组细胞在两个基板中的第一基板上的第一蛋白质供给包括第一蛋白 实验组细胞到两个基板之间的第二基板; 供给单元包括标记标签第二蛋白与第一基板和第二基板，分别当第一蛋白和第一蛋白结合分子结合到所述第一基板和第二基板，分别的细胞裂解物; 和比较和分析所述第一蛋白质和第一基板上的第二蛋白质，并在细胞裂解物的供给，以分别在第一基板和第二基板，所述第二基板之间的相互作用。 用于观察所述第一蛋白质和第二蛋白质之间的相互作用，所述第一蛋白质的每个细胞和活化水平的状态可以进行比较，并通过型细胞供给第一蛋白的变化后比较分析。

公开(公告)号：EP3153587A1

公开(公告)日：2017-04-12

申请号：EP14894116.4

申请日：2014-10-30

Applicant: Korea Advanced Institute of Science and Technology

Inventor： YOON, Tae-Young ,  LEE, Hong-Won

IPC: C12Q1/02 ,  G01N33/533 ,  G01N21/78 ,  G02B21/00

CPC classification number: G01N33/5011 ,  G01N21/6428 ,  G01N21/78 ,  G01N33/5041 ,  G01N33/543 ,  G01N33/582 ,  G01N2021/6439 ,  G01N2800/52 ,  G02B21/00

Abstract: The present invention relates to a method for analyzing the activation state of a signaling pathway in a cell or tissue separated from a subject through real time single molecule protein-protein interaction analysis, and a method for selecting a personalized medicine or predicting a therapeutic efficacy to a medicine using the same.

Abstract translation: 本发明涉及一种方法，用于在从通过实时单分子蛋白 - 蛋白相互作用分析的受试者中分离的细胞或组织分析信号传导途径的活化状态，以及用于选择个性化药物或预测治疗功效的方法 使用该药品。

公开(公告)号：EP2700947B1

公开(公告)日：2020-06-03

申请号：EP12774021.5

申请日：2012-04-20

Applicant: Korea Advanced Institute of Science and Technology

Inventor： YOON, Tae-Young

IPC: G01N33/68 ,  G01N21/64 ,  G01N33/574 ,  G01Q30/14 ,  G01Q60/20

公开(公告)号：EP2700947A2

公开(公告)日：2014-02-26

申请号：EP12774021.5

申请日：2012-04-20

Applicant: Korea Advanced Institute of Science and Technology

Inventor： YOON, Tae-Young

IPC: G01N33/533 ,  G01N21/64 ,  G01N33/68 ,  G01N33/52

Abstract: Disclosed is a method and an apparatus of analyzing protein-protein interactions at a single molecule level. The method includes binding the first proteins to the substrate where the first proteins are tagged with the first markers which bind specifically to the biomolecules immobilized on the substrate or the first proteins bind specifically to the biomolecules immobilized on the substrate; incubating the substrate bound first proteins with cell lysate containing the second proteins which are tagged with second markers; analyzing the interactions between the first proteins and the second proteins in the cell lysate, and obtaining the first analytic value representing the kinetic picture of the interactions; incubating the substrate bound first proteins with cell lysate mixture of a cell lysate consisting of the second markers-tagged second proteins and another cell lysate comprising other proteins including unlabelled second proteins and obtaining the second analytic value; comparing and analyzing the first and the second analytic values. The method and apparatus enables detailed analysis of protein-protein interactions at the single molecule level in a lysate that is equivalent to the normal intracellular environment. Also the method and apparatus enables assessment of the degrees of effects that other proteins may have on specific protein-protein interactions in the context of the normal intracellular environment.

Abstract translation: 公开的是一种方法和在单分子水平上分析蛋白质 - 蛋白质相互作用的装置。 该方法包括第一蛋白结合，其中所述第一蛋白质标记特异性结合到固定化在基片上或所述第一蛋白质生物分子特异性结合固定化在基片上生物分子的第一标记的底物; 孵育绑定细胞裂解物含有标记有第二标记的第二蛋白质第一蛋白的底物; 分析所述第一和蛋白质在细胞裂解物的第二蛋白，和获得表示的相互作用的动力学图片第一分析值之间的相互作用; 孵育绑定了细胞裂解物由第二标记标记的第二蛋白质和另一种细胞裂解物，其包括其它蛋白质，包括未标记的第二蛋白质和获得第二解析值的细胞裂解物混合物第一蛋白的底物; 比较和分析所述第一和第二分析值。 该方法和装置允许在一个裂解物的单分子水平蛋白质 - 蛋白质相互作用的详细分析所做的是相当于正常细胞内环境。 因此，该方法和设备启用程度的影响进行评估并其他蛋白质可能对正常细胞内环境的上下文中的具体蛋白质 - 蛋白质相互作用。