Method for analyzing activation state of signaling pathway and method for selecting personalized medicine using same

Patent metadata

Field	Value
Publication number	US-10845359-B2
Application number	US-201415321813-A
Country	US
Kind code	B2
Filing date	Oct 30, 2014
Priority date	Jun 3, 2014
Publication date	Nov 24, 2020
Grant date	Nov 24, 2020

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.

First claim

The invention claimed is:

1. A method of selecting a human subject for administration of an anticancer agent, comprising:
(i) determining an activation state of a signaling pathway in a cancer cell or cancer tissue separated from a human subject by (a) immobilizing a first protein in the signaling pathway on a substrate by treating the substrate with an extract of cells or tissues separated from the subject including the first protein; (b) inducing an interaction between the first protein and a second fluorescence-labeled protein by supplying to the substrate the second fluorescence-labeled protein that interacts with the first protein; and (c) analyzing interaction between the first protein and the second protein based on a fluorescent signal of a fluorescent label of the second protein, and an activation state of the signaling pathway using a total internal reflection fluorescence microscope,
(ii) selecting the subject for administration of an anticancer agent if the cell or tissue is determined to be in the activation state compared to a normal cell or tissue; and
(iii) administering the anticancer agent to the selected subject,
wherein in step (i)(c), one image is generated by averaging a predetermined number of frames and the number of single-molecule signals is measured by analyzing the image,
wherein the predetermined number of frames is obtained according to a duration of binding of the two proteins (τ on ) relative to an exposure time required to measure the fluorescent signal,
wherein the signaling pathway is one selected from EGFR, HER2, HER3 and HGFR pathway,
wherein the anticancer agent is capable of targeting EGFR, HER2, HER3 or HGFR signaling pathway,
wherein the first protein is a receptor tyrosine kinase, a Toll-like receptor, or a G protein-coupled receptor,
wherein the plurality of the second proteins is a downstream protein of the first protein.

2. The method of claim 1 , wherein the fluorescent signal analysis in the step (c) is performed by measuring the fluorescent signal in real time.

3. The method of claim 1 , further comprising
(d) repeating steps (b) and (c) using a fluorescence-labeled protein that interacts with the first protein but different from the second protein so as to analyze interactions between the first protein and a plurality of the second proteins.

Assignees

• Korea Advanced Inst Sci & Tech

Inventors

• Yoon Tae-Young
• Lee Hong Won

Classifications

• G01N33/5041

  involving analysis of members of signalling pathways · CPC title

• G01N21/78

  producing a change of colour · CPC title

• G02B21/00

  Microscopes · CPC title

• G01N2021/6439

  with indicators, stains, dyes, tags, labels, marks · CPC title

• G01N33/582

  with fluorescent label · CPC title