Exosome analysis method, exosome analysis apparatus, antibody-exosome complex, and exosome electrophoresis chip

Patent metadata

Field	Value
Publication number	US-9952215-B2
Application number	US-201514629028-A
Country	US
Kind code	B2
Filing date	Feb 23, 2015
Priority date	Aug 24, 2012
Publication date	Apr 24, 2018
Grant date	Apr 24, 2018

Abstract

A method of analyzing an exosome is provided for detecting abnormality in a cell, including: (a) a step of preparing an exosome from a sample; (b) a step of bringing the exosome prepared in the step (a) into contact with a first antibody to a protein which exists on the surface of the exosome as an antigen and forming a first antibody-exosome complex; and (c) a step of measuring a zeta potential of the first antibody-exosome complex.

First claim

What is claimed is:

1. A method of analyzing an exosome, comprising:
(a) preparing an exosome from a sample;
(b) bringing the exosome prepared in (a) into contact with a first antibody to a protein that exists on the surface of the exosome as an antigen thereby forming a first antibody-exosome complex;
(c) irradiating the first antibody-exosome complex with a laser beam to obtain a particle image of the first antibody-exosome complex using scattered light, while separating the first antibody-exosome complex from other substances through electrophoresis;
(d) determining an electrophoresis mobility of the first antibody-exosome complex based on the particle image; and
(e) determining a zeta potential of the first antibody-exosome complex based on the electrophoresis mobility of the first antibody-exosome complex.

2. The method of analyzing an exosome according to claim 1 , wherein the protein is highly expressed in an abnormal cell or a normal cell.

3. The method of analyzing an exosome according to claim 1 , wherein the protein is specifically expressed in an abnormal cell or in a normal cell.

4. The method of analyzing an exosome according to claim 1 , wherein the first antibody is formed of a plurality of types of antibodies.

5. A method of analyzing an exosome comprising;
(a) preparing an exosome from a sample;
(b) bringing the exosome prepared in (a) into contact with a first antibody to a protein that exists on the surface of the exosome as an antigen and forming a first antibody-exosome complex;
(c) measuring a zeta potential of the first antibody-exosome complex (d) bringing the exosome prepared in (a) into contact with a second antibody to a protein that exists on the surface of the exosome as an antigen and forming a second antibody-exosome complex; and
(e) measuring a zeta potential of the second antibody-exosome complex, after (c).

6. The method of analyzing an exosome according to claim 5 , wherein (e) comprises separating the second antibody-exosome complex from other substances through electrophoresis and measuring the zeta potential of the second antibody-exosome complex at one particle level.

7. The method of analyzing an exosome according to claim 6 , wherein (e) comprises measuring the zeta potential of the second antibody-exosome complex based on mobility which is optically measured.

8. The method of analyzing an exosome according to claim 6 , wherein (e) comprises measuring the zeta potential of the second antibody-exosome complex based on mobility which is measured by obtaining a particle image using scattered light.

9. The method of analyzing an exosome according to claim 5 , wherein the protein is highly and specifically expressed in an organ as an antigen.

10. An exosome analysis apparatus, comprising:
a preparation member configured for preparing an exosome from a sample;
a complex formation member configured for bringing the exosome into contact with an antibody to a protein that exists on the surface of the exosome as an antigen and thereby forming an antibody-exosome complex;
an electrophoresis member configured for separating the antibody-exosome complex from other substances through electrophoresis;
a laser beam source for irradiating the antibody-exosome complex;
a high-sensitivity camera for obtaining a particle image of the antibody-exosome complex using scattered light to determine the electrophoresis mobility of the first antibody-exosome complex; and
a zeta potential measuring member for measuring zeta potential based on the electrophoresis mobility of the first antibody-exosome complex.

11. A method of analyzing an exosome, comprising:
(a) preparing an exosome from a sample;
(b) bringing the exosome prepared in (a) into contact with a molecule that specifically binds to a biomolecule existing on the surface of the exosome thereby forming a complex;
(c) irradiating the complex with a laser beam to obtain a particle image of the complex using scattered light, while separating the complex from other substances through electrophoresis;
(d) determining an electrophoresis mobility of the complex based on the particle image; and
(e) determining a zeta potential of the complex based on the electrophoresis mobility of the first antibody-exosome complex.

12. An exosome analysis apparatus, comprising:
a preparation member configured for preparing an exosome from a sample;
a complex formation member configured for bringing the exosome into contact with a molecule that specifically binds to a biomolecule existing on the surface of the exosome to form a complex;
an electrophoresis member configured for separating the complex from other substances through electrophoresis;
a laser beam source for irradiating the complex;
a high-sensitivity camera for obtaining a particle image of the complex using scattered light to determine the electrophoresis mobility of the complex; and
a zeta potential measuring member for measuring zeta potential based on the electrophoresis mobility of the complex.

Assignees

• Univ Tokyo
• Nikon Corp

Inventors

• Ichiki Takanori
• Akagi Takanori
• Shiono Hirofumi

Classifications

• G01N33/5758 Primary

  involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites · CPC title

• G01N27/44791

  Microapparatus (sample containers with integrated microfluidic structures B01L3/5027) · CPC title

• G01N33/566 Primary

  using specific carrier or receptor proteins as ligand binding reagents {where possible specific carrier or receptor proteins are classified with their target compounds} · CPC title

• G01N33/54346

  Nanoparticles · CPC title

• G01N33/561

  Immunoelectrophoresis · CPC title