Specific coupling reaction measuring method and reagent kit and specific coupling reaction measuring apparatus for use in the same

• US 20040096909A1
• Filed: 11/12/2003
• Published: 05/20/2004
• Est. Priority Date: 11/12/2002
• Status: Active Grant

• Current Assignee: PHC Holdings Corporation
• Original Assignee: Matsushita Electric Industrial Company Limited (Panasonic Holdings Corporation)
• Inventors: Kawamura, Tatsurou, Shigetoh, Nobuyuki, Kamei, Akihito
• Granted Patent: US 7,105,359 B2
• Time in Patent Office: Days
• US Class Current: 435/7.1
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/54333 Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction

Abstract

A specific coupling reaction measuring method of this invention for measuring a content of a subject substance in a sample includes the steps of (a) constructing a reaction system including the sample and a magnetic particle on which a specific coupling substance for specifically coupling with the subject substance is immobilized; (b) measuring an optical characteristic of the reaction system; and (c) removing, from the reaction system, an agglutination complex including the subject substance, the specific coupling substance and the magnetic particle by utilizing magnetic force.

Claims

1. A specific coupling reaction measuring method for measuring a content of a subject substance in a sample, comprising the steps of:

(a) constructing a reaction system including said sample and a magnetic particle on which a specific coupling substance for specifically coupling with said subject substance is immobilized;

(b) measuring an optical characteristic of said reaction system; and

(c) removing, from said reaction system, an agglutination complex including said subject substance, said specific coupling substance and said magnetic particle by utilizing magnetic force.

2. The specific coupling reaction measuring method of claim 1,

wherein said optical characteristic is the intensity of scattered light or the quantity of transmitted light.

3. The specific coupling reaction measuring method of claim 1,

wherein said magnetic particle has a diameter of approximately 0.05 through 2 μ

m.

4. The specific coupling reaction measuring method of claim 1,

wherein said magnetic particle remaining in said reaction system is removed from said reaction system in the step (c).

5. A specific coupling reaction measuring method for measuring contents of a first subject substance and a second subject substance in a sample, comprising the steps of:

(a) constructing a reaction system including said sample and a magnetic particle on which a first specific coupling substance for specifically coupling with said first subject substance is immobilized;

(b) measuring an optical characteristic of said reaction system after the step (a);

(c) collecting an agglutination complex including said first subject substance, said first specific coupling substance and said magnetic particle by utilizing magnetic force;

(d) adding, to said reaction system, a second specific coupling substance for specifically coupling with said second subject substance; and

(e) measuring an optical characteristic of said reaction system after the step (d).

6. The specific coupling reaction measuring method of claim 5,

wherein said reaction system includes 2 through 6 wt % of polyethylene glycol in the step (d).

7. The specific coupling reaction measuring method of claim 5,

wherein said second specific coupling substance is composed of an antigen or antibody for specifically coupling with said second subject substance and a nonmagnetic particle on which said antigen or antibody is immobilized.

8. The specific coupling reaction measuring method of claim 7,

wherein said magnetic particle has a diameter of approximately 0.05 through 2 μ

m and said nonmagnetic particle has a diameter of approximately 0.05 through 2 μ

m.

9. The specific coupling reaction measuring method of claim 5,

wherein a combination of said first subject substance and said second subject substance is a combination of human hemoglobin and human albumin.

10. The specific coupling reaction measuring method of claim 5,

wherein said magnetic particle remaining in said reaction system is collected in the step (c).

11. A specific coupling reaction measuring method for measuring contents of n kinds (wherein n is an integer of 2 or more) of subject substances in a sample, comprising the steps of:

(a) constructing a reaction system including said sample and a magnetic particle on which a specific coupling substance for specifically coupling with one subject substance out of subject substances not measured yet is immobilized;

(b) measuring an optical characteristic of said reaction system after the step (a);

(c) removing, from said reaction system, an agglutination complex including said one subject substance, said specific coupling substance and said magnetic particle by utilizing magnetic force;

(d) determining to return to the step (a) when the step (b) has not been repeated (n−

1) times;

(e) adding, to said reaction system, a specific coupling substance for specifically coupling with one remaining subject substance not measured yet in said reaction system; and

(f) measuring an optical characteristic of said reaction system after the step (e).

12. The specific coupling reaction measuring method of claim 11,

wherein said magnetic particle remaining said reaction system is removed from said reaction system in the step (c).

13. A specific coupling reaction measuring method for measuring contents of n kinds (wherein n is an integer of 2 or more) of subject substances in a sample, comprising the steps of:

(a) constructing a reaction system including said sample and a specific coupling substance for specifically coupling with one subject substance out of subject substances not measured yet;

(b) measuring an optical characteristic of said reaction system after the step (a);

(c) adding, to said reaction system, a magnetic complex including a magnetic particle and a substance capable of coupling with an agglutination complex including said one subject substance and said specific coupling substance;

(d) removing, by utilizing magnetic force, said agglutination complex coupled with said magnetic complex from said reaction system;

(e) determining, after the step (d), to return to the step (a) when the step (b) has not been repeated (n−

1) times;

(f) adding, to said reaction system, a specific coupling substance for specifically coupling with one remaining subject substance not measured yet in said reaction system; and

(g) measuring an optical characteristic of said reaction system after the step (f).

14. The specific coupling reaction measuring method of claim 13,

wherein said magnetic complex not coupled with said agglutination complex is removed from said reaction system in the step (d).

15. A reagent kit for measuring contents of a first subject substance and a second subject substance in a sample, comprising:

a magnetic particle on which a first specific coupling substance for specifically coupling with said first subject substance is immobilized; and

a second specific coupling substance for specifically coupling with said second subject substance.

16. The reagent kit of claim 15,

wherein said magnetic particle has a diameter of approximately 0.05 through 2 μ

m.

17. The reagent kit of claim 15,

wherein said second specific coupling substance is composed of an antigen or antibody for specifically coupling with said second subject substance and a nonmagnetic particle on which said antigen or antibody is immobilized.

18. A reagent kit for measuring contents of n kinds (wherein n is an integer of 2 or more) of subject substances in a sample, comprising:

one specific coupling substance for specifically coupling with one subject substance out of said n kinds of subject substances; and

(n−

1) kinds of magnetic particles on which specific coupling substances for respectively specifically coupling with the other (n−

1) kinds of subject substances out of said n kinds of subject substances are respectively immobilized.

19. A reagent kit for measuring contents of n kinds (wherein n is an integer of 2 or more) of subject substances in a sample, comprising

n kinds of specific coupling substances for respectively specifically coupling with said n kinds of subject substances;

and

magnetic complexes each including a substance capable of coupling with an agglutination complex including a combination of each of (n−

1) kinds of subject substances out of said n kinds of subject substances and each of (n−

1) kinds of corresponding specific coupling substances, and a magnetic particle on which said substance is immobilized.

20. A specific coupling reaction measuring apparatus comprising:

a cell;

a light source for emitting light to said cell;

a photodetector for detecting scattered light or transmitted light received from a reaction system constructed in said cell; and

removing means for removing a magnetic particle from said cell by utilizing magnetic force when said cell contains said magnetic particle.