Biomolecule measurement apparatus

Patent metadata

Field	Value
Publication number	US-10753922-B2
Application number	US-201615779931-A
Country	US
Kind code	B2
Filing date	Nov 29, 2016
Priority date	Dec 17, 2015
Publication date	Aug 25, 2020
Grant date	Aug 25, 2020

Abstract

A biomolecule measuring device includes a first liquid tank and a second liquid tank which are filled with an electrolytic solution, a nanopore device that supports a thin film having a nanopore and is provided between the first liquid tank and the second liquid tank so as to communicate between the first liquid, tank and the second liquid tank through the nanopores, and an immobilizing member that is disposed in the first liquid tank, has a size larger than that of the thin film, and to which the biomolecules are immobilized, in which at least, one of the nanopore device and the immobilizing member has a groove structure.

First claim

The invention claimed is:

1. A biomolecule measuring device comprising:
a first liquid tank filled with an electrolyte solution;
a second liquid tank filled with the electrolytic solution;
a nanopore device that supports a thin film having a nanopore and is provided between the first liquid tank and the second liquid tank to communicate the first liquid tank with the second liquid tank through the nanopore;
an immobilizing member that is disposed in the first liquid tank, has a size larger than that of the thin film and to which biomolecules are immobilized;
a driving mechanism that drives the immobilizing member in a direction closer to or away from the thin film;
a first electrode that is provided in the first liquid tank;
a second electrode that is provided in the second liquid tank;
a stop mechanism that prevents a contact between the immobilizing member and the thin film;
a power supply that applies a voltage between the first electrode and the second electrode; and
a measurement unit that measures an ionic current flowing between the first electrode and the second electrode,
wherein at least one of the nanopore device and the immobilizing member has a groove structure in a region where the nanopore device and the immobilizing member are opposed to each other, and
wherein the measurement unit acquires sequence information on the biomolecules by an ionic current measured when the biomolecules immobilized on the immobilizing member pass through the nanopore, and
wherein the groove structure is continuously formed in a range where the nanopore device and the immobilizing member are opposed to each other.

2. The biomolecule measuring device according to claim 1 ,
wherein a cross section of the groove structure is one of a rectangle, a triangle, a semicircle, and a trapezoid.

3. The biomolecule measuring device according to claim 1 ,
wherein the biomolecules are immobilized on a convex portion of the groove structure.

4. The biomolecule measuring device according to claim 1 ,
wherein a convex portion and a concave portion of the groove structure are made of the same material.

5. The biomolecule measuring device according to claim 1 ,
wherein a convex portion and a concave portion of the groove structure are made of different materials.

6. The biomolecule measuring device according to claim 1 ,
wherein a plurality of types of biomolecules are immobilized on the immobilizing member through a plurality of different markers.

7. The biomolecule measuring device according to claim 1 ,
wherein a groove depth of the groove structure is equal to or more than 5 μm.

8. The biomolecule measuring device according to claim 1 ,
wherein a width of a convex portion of the groove structure is equal to or more than a pitch of the biomolecules immobilized on the immobilizing member.

9. The biomolecule measuring device according to claim 1 ,
wherein the driving mechanism has a function of bringing the immobilizing member into contact with the nanopore device.

10. The biomolecule measuring device according to claim 1 , further comprising:
a laser irradiation mechanism that irradiates the groove structure with a laser; and
a control unit that controls movement or rotation of the immobilizing member with the use of an image obtained by irradiation of the laser.

11. The biomolecule measuring device according to claim 1 , further comprising:
a control unit that controls movement or rotation of the immobilizing member,
wherein the control unit controls the movement or rotation of the immobilizing member by monitoring a solution resistance around the nanopore device.

12. The biomolecule measuring device according to claim 1 ,
wherein the groove structure is formed on a surface of the immobilizing member.

13. The biomolecule measuring device according to claim 1 ,
wherein the immobilizing member is made of porous silica or the porous silica is provided on a surface of the immobilizing member close to the nanopore device, and
the biomolecules to be measured are immobilized on a surface of the porous silica opposed to the nanopore device.

14. A biomolecule measuring device comprising:
a first liquid tank filled with an electrolyte solution;
a second liquid tank filled with the electrolytic solution;
a nanopore device that supports a thin film having a nanopore and is provided between the first liquid tank and the second liquid tank to communicate the first liquid tank with the second liquid tank through the nanopore;
an immobilizing member that is disposed in the first liquid tank, has a size larger than that of the thin film and to which biomolecules are immobilized;
a driving mechanism that drives the immobilizing member in a direction closer to or away from the thin film;
a first electrode that is provided in the first liquid tank;
a second electrode that is provided in the second liquid tank;
a stop mechanism that prevents a contact between the immobilizing member and the thin film;
a power supply that applies a voltage between the first electrode and the second electrode; and
a measurement unit that measures an ionic current flowing between the first electrode and the second electrode,
wherein at least one of the nanopore device and the immobilizing member has a groove structure in a region where the nanopore device and the immobilizing member are opposed to each other,
wherein the measurement unit acquires sequence information on the biomolecules by an ionic current measured when the biomolecules immobilized on the immobilizing member pass through the nanopore,
wherein the stop mechanism includes a space defining member that is disposed on the nanopore device and defines a space between the immobilizing member and the thin film, and
wherein the groove structure is formed in the space defining member.

15. A bio molecule measuring device comprising:
a first liquid tank filled with an electrolyte solution;
a second liquid tank filled with the electrolytic solution;
a nanopore device that supports a thin film having a nanopore and is provided between the first liquid tank and the second liquid tank to communicate the first liquid tank with the second liquid tank through the nanopore;
a first electrode that is provided in the first liquid tank;
a second electrode that is provided in the second liquid tank;
a power supply that applies a voltage between the first electrode and the second electrode; and
a measurement unit that measures an ionic current flowing between the first electrode and the second electrode,
wherein the first liquid tank has a micro flow path in a region in the vicinity of the nanopore,
the nanopore device has a groove structure, and
the measurement unit acquires sequence information on the biomolecules by an ionic current measured when the biomolecules pass through the nanopore.

16. A biomolecule measuring device comprising:
a first liquid tank filled with an electrolyte solution;
a plurality of second tanks filled with the electrolyte solution and electrically insulated from each other;
a nanopore device that supports a thin film having nanopores and is provided between the first liquid tank and the plurality of second tanks to communicate the first liquid tank with one of the second liquid tanks through a nanopore;
an immobilizing member that is disposed in the first liquid tank, has a size larger than that of the thin film and to which biomolecules are immobilized;
a driving mechanism that drives the immobilizing member in a direction closer to or away from the thin film;
a first electrode that is provided i

Assignees

• Hitachi High Tech Corp
• Hitachi High-Tech Corp

Inventors

• Akahori Rena
• Takeda Kenichi
• Yanagi Itaru

Classifications

• C12N15/09

  Recombinant DNA-technology · CPC title

• B01L3/502715

  characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces · CPC title

• G01N33/48721 Primary

  Investigating individual macromolecules, e.g. by translocation through nanopores (Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68) · CPC title

• G01N27/4145

  specially adapted for biomolecules, e.g. gate electrode with immobilised receptors · CPC title

• G01N27/00

  Investigating or analysing materials by the use of electric, electrochemical, or magnetic means (G01N3/00 – G01N25/00 take precedence; measurement or testing of electric or magnetic variables or of electric or magnetic properties of materials G01R) · CPC title