Method for obtaining information of test substance

• US 11,112,404 B2
• Filed: 04/12/2018
• Issued: 09/07/2021
• Est. Priority Date: 04/13/2017
• Status: Active Grant

• Current Assignee: Sysmex Corporation
• Original Assignee: Sysmex Corporation
• Inventors: Akama, Kenji, Watanabe, Toshihiro, Okada, Masaya, Yamashita, Kazuto, Ganguly, Akshay, Iwanaga, Shigeki
• Primary Examiner(s): Lam, Ann Y
• Application Number:
  US15/951,375
• Publication Number:
  US 20180299437A1
• Time in Patent Office: 1,244 Days
• Field of Search: 436528
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 2317/70 characterized by effect upon binding to a cell or to an antigen
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: G01N 2333/4709 Amyloid plaque core protein
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/5436 with ligand physically entrapped within the solid phase liposomes G01N33/5432; immunological test elements G01N33/54386
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

Disclosed is a method for obtaining information of a test substance, the method including: forming a complex by causing a capture substance to bind to a test substance in a specimen; selectively collecting at least the complex from the specimen; immobilizing the complex collected from the specimen, onto a base plate; and obtaining information regarding a structure of the test substance from the complex immobilized on the base plate.

Claims

1. A method for obtaining information of a test substance, the method comprising:

forming a complex by causing a capture substance to bind to a test substance in a specimen;

selectively collecting at least the complex from the specimen;

immobilizing the complex collected from the specimen, onto a base plate; and

obtaining information regarding a structure of the test substance from the complex immobilized on the base plate.

2. The method for obtaining the information of the test substance of claim 1, wherein

in the obtaining of the information regarding the structure of the test substance, at least one of size, morphology, structure, and aggregation degree of the test substance is obtained.

3. The method for obtaining the information of the test substance of claim 1, wherein

the capture substance includes a capture substance for labeling the test substance with fluorescence.

4. The method for obtaining the information of the test substance of claim 1, wherein

the capture substance includes a capture substance capable of binding to a solid phase.

5. The method for obtaining the information of the test substance of claim 1, wherein

the capture substance includes a capture substance capable of binding to the base plate.

6. The method for obtaining the information of the test substance of claim 3, wherein

the capture substance for labeling the test substance with fluorescence includes an antibody labeled with a fluorescent dye;

and

in the forming of the complex, the antibody labeled with the fluorescent dye is caused to bind to the test substance.

7. The method for obtaining the information of the test substance of claim 4, wherein

after the solid phase is caused to bind to the complex through the capture substance capable of binding to the solid phase, the solid phase is selectively separated in the collecting of the complex from the specimen.

8. The method for obtaining the information of the test substance of claim 7, wherein

in the collecting of the complex from the specimen, the complex is caused to be detached from the solid phase after the solid phase is selectively separated.

9. The method for obtaining the information of the test substance of claim 7, wherein

the solid phase includes a magnetic particle, and

the solid phase is selectively separated by attracting the magnetic particle by magnetic force.

10. The method for obtaining the information of the test substance of claim 7, wherein

the capture substance capable of binding to the solid phase includes an antibody which binds to the test substance and a second binding substance which binds to the solid phase,

the solid phase includes a second binding partner which specifically binds to the second binding substance, and

binding between the complex and the solid phase is realized by binding between the test substance and the antibody of the capture substance capable of binding to the solid phase and by specific binding between the second binding substance and the second binding partner.

11. The method for obtaining the information of the test substance of claim 1, wherein

the capture substance includes a capture substance capable of binding to the base plate and a capture substance capable of binding to a solid phase, and

the capture substance capable of binding to the base plate and the capture substance capable of binding to the solid phase are different from each other.

12. The method for obtaining the information of the test substance of claim 1, wherein

in the collecting of the complex from the specimen, the complex is separated from an impurity on the basis of at least one of a difference in specific gravity, a difference in size, a difference in electrical property, and a difference in immunoreaction between the complex and the impurity.

13. The method for obtaining the information of the test substance of claim 5, wherein

the capture substance capable of binding to the base plate includes an antibody which binds to the test substance and a binding substance which binds to the base plate,

the base plate includes a binding partner which specifically binds to the binding substance, and

after the antibody included in the capture substance capable of binding to the base plate is caused to bind to the test substance, the complex is immobilized on the base plate through specific binding between the binding partner and the binding substance, in the immobilizing of the complex onto the base plate.

14. The method for obtaining the information of the test substance of claim 1, wherein

after the collecting of the complex from the specimen, a capture substance capable of binding to the base plate is caused to bind to the test substance.

15. The method for obtaining the information of the test substance of claim 1, wherein

in the obtaining of the information regarding the structure of the test substance, an image of the test substance is obtained by performing image capturing of the test substance on the base plate.

16. The method for obtaining the information of the test substance of claim 1, wherein

in the collecting of the complex from the specimen, the test substance and an impurity are separated from each other, and

the test substance is a protein as a test target contained in the specimen and the impurity includes a protein other than the protein as the test target.

17. The method for obtaining the information of the test substance of claim 1, wherein

the test substance is amyloid β

.

18. The method for obtaining the information of the test substance of claim 1, wherein

the obtaining of the information regarding the structure of the test substance is performed by means of a microscope having a resolution exceeding a diffraction limit of light.

19. A method for obtaining information of a test substance, the method comprising:

causing a magnetic particle to bind to a test substance in a specimen;

selectively collecting at least the test substance from the specimen by use of the magnetic particle bound to the test substance;

immobilizing the test substance collected from the specimen, onto a base plate; and

obtaining information regarding a structure of the test substance from the test substance immobilized on the base plate.

20. A method for obtaining information of a test substance, the method comprising:

forming a complex by causing a capture substance including a fluorescent dye to bind to a test substance in a specimen;

selectively collecting at least the complex from the specimen;

immobilizing the complex collected from the specimen, onto a base plate; and

obtaining information regarding a structure of the test substance on the basis of bright spots corresponding to fluorescence emitted from a plurality of fluorescent dyes each bound to the complex immobilized on the base plate.