Fluidic device, exosome analysis method, biomolecule analysis method, and biomolecule detection method

Patent metadata

Field	Value
Publication number	US-10301682-B2
Application number	US-201615080266-A
Country	US
Kind code	B2
Filing date	Mar 24, 2016
Priority date	Sep 25, 2013
Publication date	May 28, 2019
Grant date	May 28, 2019

Abstract

The present invention provides a fluidic device, an exosome analysis method, a biomolecule analysis method, and a biomolecule detection method, which can analyze even the content of an exosome in a series of flows by introducing a sample into the device. A fluidic device of the present invention is a fluidic device which detects a biomolecule contained in an exosome in a sample, and includes: an exosome purification unit which has a layer modified with a compound having a hydrophobic chain and a hydrophilic chain; a biomolecule purification unit; a biomolecule detection unit; a first flow path which connects the exosome purification unit to the biomolecule purification unit; and a second flow path which connects the biomolecule purification unit to the biomolecule detection unit.

First claim

What is claimed is:

1. A fluidic device for detecting a biomolecule contained in an exosome in a sample, comprising:
an exosome purification unit having an exosome immobilization unit which has a layer modified with a compound having a hydrophobic chain and a hydrophilic chain, wherein the compound is configured to bind to the exosome;
a biomolecule purification unit;
a biomolecule detection unit;
a first flow path which connects the exosome purification unit to the biomolecule purification unit; and
a second flow path which connects the biomolecule purification unit to the biomolecule detection unit;
wherein the biomolecule is miRNA;
wherein the biomolecule detection unit comprises:
a substrate to which a capture probe including a sequence that can be hybridized with the first section is immobilized in a case where a target miRNA comprises a first section and a second section; and
a detection probe introduction inlet;
the detection probe having:
two stem sections forming a double stranded structure,
a loop section which is a region between the two stem sections and being labeled using a labeling substance, and
a 5′-protruding terminus or a 3′-protruding terminus, which has a sequence that can be hybridized with the second section of the target miRNA, and is protruding when the two stem sections form a double chain.

2. The fluidic device according to claim 1 ,
wherein immobilization and disruption of the exosome is performed in the exosome purification unit.

3. The fluidic device according to claim 2 ,
wherein a biomolecule present on the surface of the exosome is detected in the exosome purification unit after the immobilization of the exosome thereto.

4. The fluidic device according to claim 1 ,
wherein the exosome purification unit comprises:
an inlet.

5. The fluidic device according to claim 4 ,
wherein the exosome purification unit comprises
a sample introduction inlet and an exosome-disrupting liquid introduction inlet.

6. The fluidic device according to claim 4 ,
wherein the exosome purification unit comprises a washing solution introduction inlet.

7. The fluidic device according to claim 1 ,
wherein a ceiling height of the exosome immobilization unit is smaller than a ceiling height of the first flow path.

8. The fluidic device according to claim 1 ,
wherein the first flow path is a flow path which sends the exosome-disrupting liquid containing the biomolecule to the biomolecule purification unit from the exosome purification unit, and
wherein the second flow path is a flow path which sends a solution containing the purified biomolecule to the biomolecule detection unit.

9. The fluidic device according to claim 1 , further comprising:
a waste liquid tank for each of the exosome purification unit, the biomolecule purification unit, and the biomolecule detection unit, each of the waste liquid tank having an inlet for suctioning;
a third flow path which connects the waste liquid tank for the exosome purification unit to the exosome purification unit;
a fourth flow path which connects the waste liquid tank for the biomolecule purification-unit to the biomolecule purification unit; and
a fifth flow path which connects the waste liquid tank for the biomolecule detection unit to the biomolecule detection unit.

10. The fluidic device according to claim 1 ,
wherein the biomolecule purification unit comprises:
a biomolecule recovery liquid introduction inlet, and
a biomolecule immobilization unit.

11. The fluidic device according to claim 1 ,
wherein the exosome purification unit comprises an introduction inlet for a detection solution which is used to detect a biomolecule on the surface of an exosome.

12. A method of analyzing an exosome comprising:
providing the fluidic device of claim 1 ; and
purifying and disrupting an exosome on the exosome purification unit of the fluidic device.

13. The method of analyzing an exosome according to claim 12 , further comprising:
purifying and detecting a biomolecule released from the disrupted exosome.

Assignees

• Univ Tokyo
• Nikon Corp

Inventors

• Ichiki Takanori
• Kobayashi Masashi
• Hayashi Ayako
• Tsuchiya Shoichi
• Akagi Takanori
• Ueno Taro
• Funatsu Takashi
• Miyamoto Kenji

Classifications

• B01L2200/0631

  Purification arrangements, e.g. solid phase extraction [SPE] · CPC title

• B01L2300/0867

  Multiple inlets and one sample wells, e.g. mixing, dilution · CPC title

• C12Q1/6886 Primary

  for cancer (immunoassay for cancer G01N33/575) · CPC title

• B01L2300/161

  Control and use of surface tension forces, e.g. hydrophobic, hydrophilic · CPC title

• B01L2300/0864

  comprising only one inlet and multiple receiving wells, e.g. for separation, splitting · CPC title