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

• US 7,105,359 B2
• Filed: 11/12/2003
• Issued: 09/12/2006
• 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
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): Yu, Melanie
• Application Number:
  US10/704,802
• Publication Number:
  US 20040096909A1
• Time in Patent Office: 1,035 Days
• Field of Search: 435/4, 435/6, 435/7.1, 435/7.2, 435/7.9, 435/7.92, 435/283.1, 435/287.2, 435/287.1, 436/526, 436/525, 436/524, 436/164, 436/518
• US Class Current: 436/526
• 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 plurality of subject substances in a sample, comprising the steps of:

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

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

(c) after the step (b), removing, from said reaction system, an agglutination complex including said subject substance, said specific coupling substance and said magnetic particles by utilizing magnetic force;

(d) after the step (c), adding to said reaction system, which includes said sample, another specific coupling substance for specifically coupling with another subject substance;

wherein said another specific coupling substance includes an antigen or antibody for specifically coupling with said another subject substance; and

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

2. The specific coupling reaction measuring method of claim 1, wherein said optical characteristic is an intensity of scattered light or a quantity of transmitted light.

3. The specific coupling reaction measuring method of claim 1, wherein each of said magnetic particles has a diameter of approximately 0.05 through 2 μ

m.

4. The specific coupling reaction measuring method of claim 1, wherein a portion of said magnetic particles are included in said agglutination complex in an amount necessary to form said agglutination complex with said subject substance and said specific coupling substance, and other magnetic particles that did not form said agglutination complex remain in said reaction system in the step (a), and said other magnetic particles remaining in said reaction system are removed from said reaction system in the step (c).

5. The specific coupling reaction measuring method of claim 1, wherein said reaction system includes 2 through 6 wt % of polyethylene glycol in the step (d).

6. The specific coupling reaction measuring method of claim 1, further comprising a nonmagnetic particle on which said antigen or antibody is immobilized.

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

8. The specific coupling reaction measuring method of claim 1, wherein a combination of said subject substance and said another subject substance is a combination of human hemoglobin and human albumin.