公开(公告)号：US20160266139A1

公开(公告)日：2016-09-15

申请号：US15162673

申请日：2016-05-24

Applicant: KOREA ADVANCED INSTITUTE OF SCIENCE AND TECHNOLOGY

Inventor： Tae-Young Yoon

IPC: G01N33/68 ,  G01N21/77

CPC classification number: G01N33/6845 ,  G01N21/6428 ,  G01N21/6458 ,  G01N21/648 ,  G01N21/6486 ,  G01N21/77 ,  G01N33/574 ,  G01N2021/7786

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: 公开了在单一分子水平分析蛋白质 - 蛋白质相互作用的方法。 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：制备至少两个底物，其中作为与第一蛋白质结合的生物分子的第一蛋白结合分子附着于每个底物; 通过将对照组细胞中包含的第一个蛋白质提供给两个底物中的第一个底物并分别提供第一个蛋白质和第一个蛋白质，分别诱导第一个蛋白质和第一个蛋白质结合分子上的第一个蛋白质结合分子之间的结合 在实验组细胞中两片基片之间的第二衬底; 当第一蛋白质和第一蛋白结合分子分别结合到第一衬底和第二衬底时，分别将第一蛋白质和第一蛋白结合分子结合到第一底物和第二底物的细胞裂解液，包括标记标记的第二蛋白质的细胞裂解液; 以及分别在第一底物和第二底物上的第一蛋白质和第二蛋白质之间的相互作用以分别将细胞裂解物供应到第一基底和第二底物。 为了观察第一种蛋白质和第二种蛋白质之间的相互作用，可以通过比较提供第一种蛋白质的细胞类型的变化来比较和分析每种细胞的状态和第一种蛋白质的活化水平。

公开(公告)号：US09377462B2

公开(公告)日：2016-06-28

申请号：US14059294

申请日：2013-10-21

Applicant: Korea Advanced Institute of Science and Technology

Inventor： Tae-Young Yoon

IPC: G01N31/00 ,  G01N33/53 ,  G01N33/574 ,  G01N33/68 ,  G01N21/64

CPC classification number: G01N33/6845 ,  G01N21/6428 ,  G01N21/6458 ,  G01N21/648 ,  G01N21/6486 ,  G01N21/77 ,  G01N33/574 ,  G01N2021/7786

Abstract: A method of analyzing protein-protein interactions includes: preparing 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; and comparing 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.

Abstract translation: 分析蛋白质 - 蛋白质相互作用的方法包括：制备两个底物，其中作为与第一蛋白质结合的生物分子的第一蛋白结合分子附着于每个底物; 通过将对照组细胞中包含的第一个蛋白质提供给两个底物中的第一个底物并分别提供第一个蛋白质和第一个蛋白质，分别诱导第一个蛋白质和第一个蛋白质结合分子上的第一个蛋白质结合分子之间的结合 在实验组细胞中两片基片之间的第二衬底; 将包括标记标记的第二蛋白的细胞的细胞裂解物分别提供给第一底物和第二底物; 以及分别在第一底物和第二底物上的第一蛋白质和第二蛋白质之间的相互作用，将细胞裂解物供应到第一底物和第二底物。

公开(公告)号：US09423400B2

公开(公告)日：2016-08-23

申请号：US14059349

申请日：2013-10-21

Applicant: Korea Advanced Institute of Science and Technology

Inventor： Tae-Young Yoon

IPC: G01N31/00 ,  G01N33/53 ,  G01N33/574 ,  G01N33/68 ,  G01N21/64

CPC classification number: G01N33/6845 ,  G01N21/6428 ,  G01N21/6458 ,  G01N21/648 ,  G01N21/6486 ,  G01N21/77 ,  G01N33/574 ,  G01N2021/7786

Abstract: A method of analyzing protein-protein interactions 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.

Abstract translation: 分析蛋白质 - 蛋白质相互作用的方法包括将第一蛋白质与底物结合，其中第一蛋白质与第一标记物结合，第一标记物特异性结合固定在底物上的生物分子，或者第一种蛋白质特异性结合固定在底物上的生物分子; 将底物结合的第一蛋白质与含有用第二标记物标记的第二蛋白质的细胞裂解物孵育; 分析细胞裂解物中第一种蛋白质和第二种蛋白质之间的相互作用，并获得表示相互作用动力学图像的第一个分析值; 将底物结合的第一蛋白质与由第二标记标记的第二蛋白质组成的细胞裂解物和包含其他蛋白质的另一细胞裂解物（包括未标记的第二蛋白质）的细胞裂解物混合物孵育并获得第二分析值; 比较和分析第一和第二个分析值。

公开(公告)号：US20140113307A1

公开(公告)日：2014-04-24

申请号：US14059294

申请日：2013-10-21

Applicant: Korea Advanced Institute of Science and Technology

Inventor： Tae-Young Yoon

IPC: G01N33/574

CPC classification number: G01N33/6845 ,  G01N21/6428 ,  G01N21/6458 ,  G01N21/648 ,  G01N21/6486 ,  G01N21/77 ,  G01N33/574 ,  G01N2021/7786

Abstract: A method of analyzing protein-protein interactions includes: preparing 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; and comparing 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.

Abstract translation: 分析蛋白质 - 蛋白质相互作用的方法包括：制备两个底物，其中作为与第一蛋白质结合的生物分子的第一蛋白结合分子附着于每个底物; 通过将对照组细胞中包含的第一个蛋白质提供给两个底物中的第一个底物并分别提供第一个蛋白质和第一个蛋白质，分别诱导第一个蛋白质和第一个蛋白质结合分子上的第一个蛋白质结合分子之间的结合 在实验组细胞中两片基片之间的第二衬底; 将包括标记标记的第二蛋白的细胞的细胞裂解物分别提供给第一底物和第二底物; 以及分别在第一底物和第二底物上的第一蛋白质和第二蛋白质之间的相互作用，将细胞裂解物供应到第一底物和第二底物。

公开(公告)号：US10401367B2

公开(公告)日：2019-09-03

申请号：US15945890

申请日：2018-04-05

Applicant: KOREA ADVANCED INSTITUTE OF SCIENCE AND TECHNOLOGY

Inventor： Tae-Young Yoon

IPC: G01N31/00 ,  G01N33/53 ,  G01N33/68 ,  G01N21/64 ,  G01N33/574 ,  G01N21/77

Abstract: A method of analyzing protein-protein interactions 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.

公开(公告)号：US20140113309A1

公开(公告)日：2014-04-24

申请号：US14059349

申请日：2013-10-21

Applicant: Korea Advanced Institute of Science and Technology

Inventor： Tae-Young Yoon

IPC: G01N21/64

CPC classification number: G01N33/6845 ,  G01N21/6428 ,  G01N21/6458 ,  G01N21/648 ,  G01N21/6486 ,  G01N21/77 ,  G01N33/574 ,  G01N2021/7786

Abstract: A method of analyzing protein-protein interactions 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.

Abstract translation: 分析蛋白质 - 蛋白质相互作用的方法包括将第一蛋白质与底物结合，其中第一蛋白质与第一标记物结合，第一标记物特异性结合固定在底物上的生物分子，或者第一种蛋白质特异性结合固定在底物上的生物分子; 将底物结合的第一蛋白质与含有用第二标记物标记的第二蛋白质的细胞裂解物孵育; 分析细胞裂解物中第一种蛋白质和第二种蛋白质之间的相互作用，并获得表示相互作用动力学图像的第一个分析值; 将底物结合的第一蛋白质与由第二标记标记的第二蛋白质组成的细胞裂解物和包含其他蛋白质的另一细胞裂解物（包括未标记的第二蛋白质）的细胞裂解物混合物孵育并获得第二分析值; 比较和分析第一和第二个分析值。

公开(公告)号：US09964544B2

公开(公告)日：2018-05-08

申请号：US15642427

申请日：2017-07-06

Applicant: KOREA ADVANCED INSTITUTE OF SCIENCE AND TECHNOLOGY

Inventor： Tae-Young Yoon

IPC: G01N33/68 ,  G01N21/64 ,  G01N33/574 ,  G01N21/77

CPC classification number: G01N33/6845 ,  G01N21/6428 ,  G01N21/6458 ,  G01N21/648 ,  G01N21/6486 ,  G01N21/77 ,  G01N33/574 ,  G01N2021/7786

Abstract: A method of analyzing protein-protein interactions 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.

公开(公告)号：US09739785B2

公开(公告)日：2017-08-22

申请号：US15162673

申请日：2016-05-24

Applicant: KOREA ADVANCED INSTITUTE OF SCIENCE AND TECHNOLOGY

Inventor： Tae-Young Yoon

IPC: G01N31/00 ,  G01N33/53 ,  G01N33/68 ,  G01N21/64 ,  G01N33/574 ,  G01N21/77

CPC classification number: G01N33/6845 ,  G01N21/6428 ,  G01N21/6458 ,  G01N21/648 ,  G01N21/6486 ,  G01N21/77 ,  G01N33/574 ,  G01N2021/7786

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.

公开(公告)号：US09733255B2

公开(公告)日：2017-08-15

申请号：US15211096

申请日：2016-07-15

Applicant: KOREA ADVANCED INSTITUTE OF SCIENCE AND TECHNOLOGY

Inventor： Tae-Young Yoon

IPC: G01N31/00 ,  G01N33/53 ,  G01N33/68 ,  G01N21/64 ,  G01N33/574 ,  G01N21/77

CPC classification number: G01N33/6845 ,  G01N21/6428 ,  G01N21/6458 ,  G01N21/648 ,  G01N21/6486 ,  G01N21/77 ,  G01N33/574 ,  G01N2021/7786

Abstract: A method of analyzing protein-protein interactions 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.